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黄花蒿组培快繁与种质离体保存的研究 总被引:3,自引:0,他引:3
以带侧芽的黄花蒿(Artemisia annua L.)茎段为外植体,以MS为基本培养基,进行组织培养和种质保存研究.结果表明,培养基MS 6-BA 1.0 mg L-1 IBA 0.1 mg L-1、MS 6-BA 0.5 mg L-1 IBA 0.1 mg L-1和MS NAA0.1 nag L-1 IBA 0.5 rng L-1可分别用于黄花蒿的芽诱导、增殖和生根培养,培养20 d的增殖倍数为5.5倍,生根率98.3%.培养基MS CCC 1.0 mg L-1、MS CCC 2.0 nag L-1、MS PP3334.0 mg L-1可用作离体保存,连续保存200 d的存活率分别达72.3%、77.0%、69.2%.活力检测表明,黄花蒿种质经保存后的增殖、生根能力没有下降.因此,可通过诱导腋芽增殖建立黄花蒿快繁体系,及在培养基中添加CCC或PP333拼能使材料长期保存. 相似文献
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A simple TLC-densitometric technique has been developed for the rapid and accurate analysis of artemisinin in a large number of Artemisia annua plantlets cultured in vitro. This new analytical method is based on the structural conversion of artemisinin on a silica gel layer by ammonia vapour to form 10-azadesoxyartemisinin, a chromophore-containing compound (lambdamax 320 nm) that can be detected by UV-based TLC densitometry. The TLC system was evaluated quantitatively in terms of product stability, precision, accuracy and calibration. Good linearity was obtained in the range of 0.01-0.12 microg artemisinin. The technique appeared to be accurate and sensitive as compared with the complicated pre-column reaction-HPLC technique. Among 90 samples of A. annua plantlets, the artemisinin content in the leaves appeared to be highly variable, ranging from 0.02 to 0.67% w/w dry weight. These results demonstrate that densitometric TLC can be a cheap and simple technique for the accurate screening of high-artemisinin-producing plants. 相似文献
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Dedifferentiated and differentiated tissue cultures ofArtemisia annua L. for artemisinin production were carried out. The calluses were initiated on MS medium supplemented with sucrose (30 g l-1), myoinositol (100 mg l-1) and RT vitamins. The auxins used were naphtaleneacetic acid (NAA), indoleacetic acid (IAA), indolebutyric acid (IBA) and 2,4-dichlorophenoxyacetic acid (2,4-d). These were added to the basal medium either singly or in combination. The best results were obtained with 2.4-d (4.5 M : 0.02 d-1) and NAA (5.4 M : 0.06 d-1). Cell suspensions were established on the same media without agar. Suspension cultures showed different morphological characteristics according to the plant growth regulator supplied. Organized cultures were initiated from callus obtained on 2,4-d (4.5 M) and from bud cultures. Medium containing 6-benzylaminepurine (BA) (8.9 M)+NAA (0.54 M); Zeatin (45.62 M)+NAA (5.37 M) or BA (8.9 M) stimulated both organogenesis in callus (frequency of induction =50%) and semi-organized tissue in shoot buds. BA (13.32 M)+NAA (1.08 M) or BA (13.32 M) only stimulated multiple shoot cultures (frequency of induction =80%). Regarding artemisinin content, while the values obtained were 1.13 and 0.78 mg gDW-1 in primary callus, artemisinin was not detected in cell suspension and only traces of it were found in multiple shoot cultures. 相似文献
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Artemisinin, a sesquiterpene lactone isolated from the Chinese medicinal plant Artemisia annua L., is an effective antimalarial agent, especially for multi-drug resistant and cerebral malaria. To date, A. annua is still the only commercial source of artemisinin. The low concentration of artemisinin in A. annua, ranging from 0.01 to 0.8% of the plant dry weight, makes artemisinin relatively expensive and difficult to meet the demand
of over 100 million courses of artemisinin-based combinational therapies per year. Since the chemical synthesis of artemisinin
is not commercially feasible at present, another promising approach to reduce the price of artemisinin-based antimalarial
drugs is metabolic engineering of the plant to obtain a higher content of artemisinin in transgenic plants. In the past decade,
we have established an Agrobacterium-mediated transformation system of A. annua, and have successfully transferred a number of genes related to artemisinin biosynthesis into the plant. The various aspects
of these efforts are discussed in this review. 相似文献
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Cryopreservation of callus tissue of Artimisia annua L. was optimized. Two lines of calli were precultured on MS medium with 5% (v/v) dimethyl sulfoxide, and protected by a cryoprotectant containing 15% (v/v) ethylene glycol, 15% (v/v) dimethyl sulfoxide, 30% (v/v) glycerol and 13.6% (w/v) sucrose. The highest survival rate of callus A201 reached 87% after it was pretreated at 25°C, cryopreserved by liquid nitrogen, recovered in water bath at 25°C and reloaded at 25°C with 34% (w/v) sucrose solution, and that of callus A202 reached 78% after it was treated as callus A201, except pretreated at 35°C, recovered at 35°C and reloaded with 47.8% (w/v) sucrose solution. 相似文献
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Phithak Inthima Masahiro Otani Tomoya Hirano Yoriko Hayashi Tomoko Abe Masaru Nakano Kanyaratt Supaibulwatana 《Plant Cell, Tissue and Organ Culture》2014,119(1):131-139
Artemisia annua L. is a commercial source of artemisinin. Nevertheless, artemisinin content within the plant is relatively low and varies depending on genotype and environment. To broaden the genetic variability, the mutation effect of 12C-ion beam irradiation on A. annua was examined. Irradiation at 2.5 Gy had a slight lethal effect to nodal segments while a noticeable lethal effect was observed at 5 and 10 Gy. Furthermore, at higher doses (20 and 50 Gy), a severe lethal effect was observed. Mutations at the DNA level of axillary bud-derived shoots were performed by RAPD. The mutation frequency at 10 Gy was about 1.7 and 2.1 times higher than that at 2.5 and 5 Gy, respectively. After growth and artemisinin production observation of 72 irradiated mutants, around 14 and 7 % of them showed higher artemisinin content and artemisinin yield compared to the controls, respectively. The highest artemisinin content in a mutant was 1.43 % DW, which was 3.2-fold higher than the original wild type. Additionally, the highest artemisinin yield in mutants was 3.68 mg/plant, which was around 1.4-fold higher than in the wild type. Moreover, irradiated mutants exhibited antibacterial activity against S. aureus, but the wild types did not. This study presents an effective application of heavy ion beam irradiation to create variations and improve artemisinin production in A. annua. 相似文献
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本文建立了反相高效液相色谱快速测定青蒿中青蒿酸含量的方法。色谱条件为:Diamonsil C18色谱柱(250 mm×4.6 mm,5μm),柱温为(30±1)℃,流动相采用乙腈与0.2%磷酸水溶液混合液(体积比65:35),流速1 mL/min,检测波长220 nm。标准曲线回归方程:Y=8.784573×10-8X-6.443559×10-5,r=0.9997,青蒿酸回收率为102.4%。实验证明该方法稳定可靠、精密度高、重现性好、简单可行,适用于青蒿酸的分析检测。 相似文献
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Factors influencing Agrobacterium tumefaciens-mediated transformation of Artemisia annua L. 总被引:2,自引:0,他引:2
A. Vergauwe E. Van Geldre D. Inzé M. Van Montagu E. Van den Eeckhout 《Plant cell reports》1998,18(1-2):105-110
Following our previously described Agrobacterium tumefaciens-mediated transformation procedure for Artemisia annua L., we have undertaken several additional experiments to establish the importance of some parameters such as explant type,
age of explant source, A. tumefaciens strain and type of binary vector. Several binary vectors were useful for the production of transgenic callus on explants
of different ages. In transformed calli, a good correlation between integration and expression of foreign DNA was observed:
different assays showed expression of β-Glucuronidase, neomycin phosphotransferase II, superoxide dismutase and bleomycin acetyl transferase. The regeneration of
transgenic plants required more restricted conditions. Only with the pTJK136 vector could transgenic plants be obtained from
leaf and stem explants from 12- to 18-week-old plants. Co-cultivation for 48 h seemed favorable for the regeneration of transgenic
plants. Stable integration and expression of the transgenes was also shown in the progeny.
Received: 18 August 1997 / Revision received: 3 December 1997 / Accepted: 3 July 1998 相似文献
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Artemisinin, the endoperoxide sesquiterpene lactone, is an effective antimalarial drug isolated from the Chinese medicinal plant Artemisia annua L. Due to its effectiveness against multi-drug-resistant cerebral malaria, it becomes the essential components of the artemisinin-based combination therapies which are recommended by the World Health Organization as the preferred choice for malaria tropica treatments. To date, plant A. annua is still the main commercial source of artemisinin. Although semi-synthesis of artemisinin via artemisinic acid in yeast is feasible at present, another promising approach to reduce the price of artemisinin is using plant metabolic engineering to obtain a higher content of artemisinin in transgenic plants. In the past years, an Agrobacterium-mediated transformation system of A. annua has been established by which a number of genes related to artemisinin biosynthesis have been successfully transferred into A. annua plants. In this review, the progress on increasing artemisinin content in A. annua by transgenic approach and its future prospect are summarized and discussed. 相似文献
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《Environmental and Experimental Botany》2005,53(2):185-193
The Artemisia annua L. plants were subjected separately to NaCl (0–160 mM) and lead acetate (0–500 μM) at the age of 90 days (S1 treatment) and 120 days (S2 treatment). The treated plants were studied on 100, 130 and 160 days after sowing (DAS) in S1, and on 130 and 160 DAS in S2 treatments for lipid peroxidation rate, photosynthetic rate (Pn), chlorophyll content, artemisinin concentration and artemisinin yield in leaf samples and for total biomass accumulation. The treatments enhanced lipid peroxidation at all stages of plant growth and increased the concentration and yield of artemisinin at 100 and 130 DAS in S1 and S2, respectively, while other parameters declined at all growth stages. The magnitude of changes was greater in lead-treated than in salt-treated plants. Both treatments induced oxidative stress which might have damaged the photosynthetic apparatus resulting in a loss of chlorophyll content and a decline in photosynthetic rate, biomass accumulation and artemisinin production. The increase in artemisinin content, observed during the early phase of plant growth, might be due to a sudden conversion of its precursors (e.g. artemisinic acid/dihydroartemisinic acid) to artemisinin by activated oxygen species under oxidative stress. 相似文献
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T. E. Soktoeva G. L. Ryzhova K. A. Dychko V. V. Khasanov S. V. Zhigzhitzhapova L. D. Radnaeva 《Russian Journal of Bioorganic Chemistry》2013,39(7):761-764
Isolation of artemisinin from Artemisia annua L. and its quantification by the HPLC-MS method are considered. Different extraction methods were used for the isolation of artemisinin: maceration, ultrasonic and subcritical CO2-extraction. The component content of the CO2- and hexane extracts was studied by the GC-MS method. 相似文献
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Annemieke Vergauwe Ronny Cammaert Dirk Vandenberghe Christiane Genetello Dirk Inze Marc Van Montagu Elfride Van den Eeckhout 《Plant cell reports》1996,15(12):929-933
Summary A transformation system was developed for Artemisia annua L. plants. Leaf explants from in vitro grown plants developed callus and shoots on medium with 0.05 mg/L naphthaleneacetic acid and 0.5 mg/L N6-benzyladenine after transformation with the C58C1 RifR (pGV2260) (pTJK136) Agrobacterium tumefaciens strain. A concentration of 20 mg/L kanamycin was added in order to select transformed tissue. Kanamycin resistant shoots were rooted on naphthaleneacetic acid 0.1 mg/L. Polymerase chain reactions and DNA sequencing of the amplification products revealed that 75% of the regenerants contained the foreign genes. 94% of the transgenic plants showed a -glucuronidase-positive response.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BA
N6-benzyladenine
- GM
germination medium
- GMVIT
germination medium with vitamins
- GUS
-glucuronidase
- Kin
kinetin (N6-furfurylaminopurine)
- NAA
-naphthaleneacetic acid
- NPT II
neomycin phosphotransferase II
- PCR
Polymerase Chain Reaction
- T-DNA
transfer-DNA
- X-glucuronide
5-bromo-4-chloro-3-indolyl -D-glucuronide 相似文献