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用根癌农杆菌(Agrobacterium tumefaciens)菌株C58感染栝楼(Trichosanthes kirilowii Maxim.)无菌苗诱导冠瘿瘤,获得其冠瘿组织;栝楼冠瘿组织经除菌后能在无激素的MS培养基上良好生长,纸电泳检测结果表明其合成了冠瘿碱,表明Ti质粒转化成功。栝楼冠瘿组织在MS培养基上形成完整植株,移栽后良好生长。 相似文献
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商陆毛状根的诱导、培养及其扼甙的产生 总被引:9,自引:0,他引:9
发根农杆菌(Agrobacterium rhizogenes)R1601感染商陆叶片外植体1周后,在其切口处产生毛状根,20d后产生毛状根的外植体比例达70%,毛状根可直接从叶片外植体叶脉处或从叶脉处产生的愈伤组织上产生,毛状根能在无激素的MS培养基上自主生长,其呼吸速率比对照根提高85.6%,冠瘿碱检测和PCR扩增结果表明,发根农杆菌RiT-DNA的冠瘿碱合成酶基因及其Ri质粒的rol基因均已在商陆毛状根基因组中得到表达。毛状根中总皂甙含量约为自然根的1.54倍,但其多糖含量则仅为非转化根的70%。 相似文献
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商陆毛状根的诱导、培养及其皂甙的产生 总被引:1,自引:0,他引:1
发根农杆菌(Agrobacterium rhizogenes)R1601 感染商陆叶片外植体1周后,在其切口处产生毛状根,20d后产生毛状根的外植体比例达70%;毛状根可直接从叶片外植体叶脉处或从叶脉处产生的愈伤组织上产生。毛状根能在无激素的 MS培养基上自主生长,其呼吸速率比对照根提高85.6%。冠瘿碱检测和PCR扩增结果表明,发根农杆菌RiTDNA的冠瘿碱合成酶基因及其Ri质粒的rol 基因均已在商陆毛状根基因组中得到表达。毛状根中总皂甙含量约为自然根的154倍,但其多糖含量则仅为非转化根的70%。 相似文献
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丹参冠瘿组织高产株系选择和丹参酮的产生 总被引:14,自引:0,他引:14
丹参(Salvia miltiorrhiza)是治疗心血管系统疾病的重要中草药。前文我们已报道了丹参冠瘿组织培养的部分研究结果[1].利用根癌农杆菌(Agrobacterium tumefaciens)Ti 质粒转化产生的冠瘿组织进行高产株系选择的研究还很少,本文报道丹参冠瘿组织高产株系选择和丹参酮产生的研究结果。 相似文献
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农杆菌T—DNA介导的植物转基因的分子机制 总被引:4,自引:0,他引:4
前言 根瘤农杆菌(Agrobacterium tumefaciens)与发根农根菌(A.rhizogenes)同属根瘤菌科,是革兰氏阴性植物病原菌。前者感染植物引起冠瘿病(Crop gall tumour),冠瘿病用含有抗生素的培养基培养与除菌。可无限增殖;后者感染植物诱发出许多不定根,把不定根培养在上述培养基上也可迅速生长,多次分枝成毛状称毛状根(hairy root)。农杆菌感染机理很复杂。其感染作用起始于受伤的植物细胞分泌的大量化学物质时期。受伤植物细胞分泌的酚分子诱导农杆菌怀有的Ti质粒或Ri质粒上的基因活化,尔后农杆菌紧附植物细胞并把Ti质粒或Ri质粒上一部分DNA(称作T—DNA,tranfer DNA)转移到植物染色体上。T.DNA共价整合后编译合成的新的低分量的代谢物,称为冠瘿碱。 相似文献
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本文利用发根农杆菌感染黄瓜植体得到的转化根,在不同激素配比的分分培养基上培养,进行再生植株的诱导试验,结果表明,NAA和BA的合适配比,有利于黄瓜植株的再生,转化根及其再生植株检测到冠瘿碱的存在,说明农杆菌Ri质粒的T-DNA已转移到黄瓜基因组中。 相似文献
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本文报道了致瘤农杆菌(Agrobacterium lume/aciens)的T(?)菌株对分属于五大类型的41个品种的栽培种花生致瘤实验结果。在41个品种中的6个品种上诱发了畸胎瘤。在无激素的MS培养基上诱发冠瘿瘤产生了愈伤组织。瘤组织、愈伤组织,畸胎瘤上幼苗的茎和叶均含有胭脂碱。 相似文献
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M Krsnik-Rasol 《The International journal of developmental biology》1991,35(3):259-263
Peroxidase was studied as a developmental marker in pumpkin (Cucurbita pepo L.) callus lines and horse-radish (Armoracia lapathifolia Gilib) transformants. Embryogenic callus lines DE grown on MS medium with 2.4-D and NA-3 grown on medium with NAA and adenine sulfate showed about a 20 times higher enzyme activity than the habituated non-embryogenic line Z5b/T grown on medium without hormones. A rise in peroxidase activity indicated that somatic embryogenesis was triggered in a few habituated tissue cultures. Separated globular embryoids had a manifold lower enzyme activity than the callus from which they originated. SDS-electrophoresis showed distinct polypeptide patterns between the horse-radish leaves and crown galls, but the tumor characteristic protein bands failed to be identified. In horse-radish crown galls and short bushy plants regenerated from hairy roots an enhanced peroxidase activity was registered. Due to its high peroxidase level and abundant biomass production horse-radish transformants should facilitate enzyme production. 相似文献
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Awadhesh Kumar Arya Kottackal Poulose Martin Aneta Sabovljevic Joseph Madassery 《Acta Physiologiae Plantarum》2011,33(5):2011-2017
A simple and easy transformation strategy was accomplished on field growing plants of Phyllanthus amarus, an anti-hepatitis B drug plant. Infection of Agrobacterium rhizogenes strains A4M70GUS and ATCC 15834 on decapitated shoots of field growing P. amarus induced hairy roots and crown gall, respectively. Infection with A4M70GUS yielded a mean of 23.2 roots from 40% plants in 40-day period. The crown gall induced on 30% plants after infection
with ATCC 15834 grew to 5–10 mm in diameter. The roots and crown galls established in vitro on Murashige and Skoog (MS) basal
medium grew well. The hairy roots yielded fivefold (6.91 g) biomass in half-strength MS liquid medium to that of the adventitious
roots derived from internode explants in MS medium with 8.0 μM α-naphthaleneacetic acid (1.39 g). Histochemical assay and
PCR analysis using the primers of uidA coding region confirmed the hairy roots induced by A4M70GUS. The crown galls induced by ATCC 15834 were confirmed by PCR analysis using rolB gene primers. The protocol enables an easy and early accomplishment of hairy roots. 相似文献
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Ooms G. Karp A. Roberts J. 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1983,64(2):169-175
Theoretical and Applied Genetics - Agrobacterium tumefaciens strains, known to induce tobacco crown galls that spontaneously develop shoots, were used to induce galls on cultured shoots of a... 相似文献
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Pelet , F., A. C. Hildebrandt , A. J. Riker, and F. Skoog . (U. Wisconsin, Madison.) Growth in vitro of tissues isolated from normal stems and insect galls . Amer. Jour. Bot. 47(3) : 186—195. Illus. 1960.–In a preliminary analysis of the nature of gall formation induced by insects, a comparative study has been made of the in vitro growth and nutrition of plant tissues derived from insect galls and from normal plants. Grape, elm, poplar, and willow tissues were grown on a standard medium, modified White's nutrient medium, with coconut milk and/or various growth factors added. Satisfactory growth was obtained over a temperature range from 16° to 36°C. but was generally optimal at 28°—32°C. The optimum pH was generally 4.0—4.5, but a pH of 6.0 or 7.0 gave better growth when the medium contained 2,4-dichlorophenoxyacetic acid. Detailed nutritional studies were limited to grape tissue. Excised stems and excised galls induced by Phylloxera vastatrix Planch, were grown on the basal medium with vitamins and supplemented with naphthaleneacetic acid, indoleacetic acid, kinetin, casein hydrolysate, yeast extract, adenine and a few amino acids added in various combinations. Growth (fresh weight) was measured after a 6-week growth period. When these substances were added singly the optimal concentrations and the quality of growth of stem explants were as follows: with adenine (40 mg./l.) or kinetin (1 mg./l.), growth poor; with NAA (1 mg./l.) or IAA (2 mg./l.), growth fair; and with the only concentration of a powdered casein hydrolysate (3 g./l.), growth good. Gall explants responded more readily to kinetin or adenine but did not form callus in the presence of casein hydrolysate alone. Stem tissues formed both roots and callus, whereas gall tissues formed only callus. The same substances were tested in various combinations. NAA and kinetin provided for moderate, continuous growth, and excellent growth if casein hydrolysate and adenine also were added to the medium. The NAA requirements were markedly reduced in the grape tissues which had been subcultured for 1 or 4 years on coconut milk medium. Friable tissue types were inhibited by the adenine and casein hydrolysate combinations. They grew through 1 passage only on basal medium and then died if not supplied with NAA and kinetin. Firm tissues responded favorably, although irregularly, to casein hydrolysate and adenine. It was concluded that although nutrient requirements varied with tissues derived from insect galls and from normal plants, they also varied with the time of cultivation in vitro. The induction of galls by Phylloxera was not a permanent change in growth factor requirements comparable to that conferred by the crown gall bacteria. In attempts to grow the insect in sterile culture in vitro 5 successive generations of phylloxera were reared on callus tissue. 相似文献
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Methylglyoxal (MG) is a highly reactive α-oxoaldehyde, demonstrating anticancer effect on plant neoplastic tumours. In in vivo studies it was observed that MG destroyed crown gall tumours in Nicotiana tabacum produced by Agrobacterium tumefaciens, without any adverse effect on the host. The efficacy of MG in comparison to other anticancer drugs viz. cisplatin and ellagic acid in the treatment of crown gall was investigated. A slight degeneration of galls was noted in
plants treated with cisplatin and ellagic acid but the plants died subsequently. With MG however, crown galls were completely
cured and the plants completed their usual life cycle by flowering and producing seeds. MG inhibited the respiration of crown
gall calluses suggesting that energy depletion resulted in tumour destruction. 相似文献
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Selective media for three biovars of Agrobacterium 总被引:3,自引:1,他引:2
Three selective media for the isolation of the three known biovars of Agrobacterium are described. Selectivity was based on carbon and nitrogen sources; L(—)-arabitol for biovar 1, erythritol for biovar 2 and a combination of tartrate and D-glutamate for biovar 3. The new media were compared with existing selective media. Recovery of agrobacteria from soil was very efficient and discrimination between the three biovars was satisfactory except for tartrate-utilising biovar 1 isolates which grew on the biovar 3 medium. Some strains of Pseudomonas sp. isolated from crown galls on vine could utilise octopine as a source of carbon and nitrogen. 相似文献