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1.
Overexpression of mtlD gene in transgenic Populus tomentosa improves salt tolerance through accumulation of mannitol 总被引：2，自引：0，他引：2
The mtlD gene encoding mannitol-1-phosphate dehydrogenase, which catalyzes the biosynthesis of mannitol from fructose, was cloned from Escherichia coli and transferred to poplar (Populus tomentosa Carr.) through Agrobacterium-mediated transformation. The transgenic plants were screened and selected on Murashige and Skoog (MS) medium containing 30-50 mg l(-1) kanamycin and verified by polymerase chain reaction (PCR) and Southern blotting. Expression of the gene led to synthesis and accumulation of mannitol in the transgenic plants. Gas chromatography and mass spectrometry (GC/MS) and capillary gas chromatography (GC) showed that transgenic plants accumulated much more mannitol in their tissues than the wild-type plants, whether cultured in vitro, or grown hydroponically or in the field. Increased salt tolerance of transgenic plants was observed both in vitro and in hydroponic culture. The transgenic buds rooted normally on MS medium containing 50 mM NaCl, whereas wild-type buds did not. In the 40-day hydroponic experiments, transgenic poplar plants survived in a 75-mM NaCl treatment, whereas the wild-type poplar plants tolerated only 25 mM NaCl. Under the same NaCl stress, stomatal conductance, transpiration rates and photosynthetic rates were all higher in transgenic plants than in wild-type plants, whereas cellular relative conductivity was lower. We demonstrated that the mtlD gene was expressed in transgenic poplar plants, resulting either directly or indirectly in mannitol accumulation and improved salt tolerance. The constant mannitol concentrations in transgenic plants during the NaCl treatments indicated that mannitol accumulation caused by the mtlD gene was not a consequence of NaCl stress. Height growth was reduced by about 50% in the transgenic plants compared with the wild-type plants in the absence of salt; however, relative growth rate was much less influenced by salt stress in transgenic plants than in wild-type plants. The stunted growth of the transgenic plants may in part explain their improved salt tolerance. 相似文献
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爬行卫矛下胚轴高频离体再生体系的建立(英文) 总被引：1，自引：0，他引：1
In this paper,a protocol for efficient shoot regeneration was successfully developed from hypocotyl explants of Euonymus fortunei var.radicans.Some factors that influenced shoot regeneration such as different combinations of plant growth regulators,types of medium and inoculation ways were studied in order to establish an efficient plant regeneration for transformation.The results showed that hypocotyl explants wero horizontally cultured on a basic medium composed of MS medium supplemented with 0.5 mg·L-1 BAP and 0.01 mg·L-1 NAA for induction and development of adventidous shoots.Ninety-four percent of regeneration frequency and 5.1 shoots per explants were obtmned after 30 days of culture.Regenerated shootsproliferated efficiently on a shoot multiplication medium consisting of MS medium containing 1.0 mg·L-1 BAP and 0.1 mg·L-1 NAA.Microshoots were rooted on a rooting medium made up of MS medium enriched with O.5 mg·L-1 IBA and O.5 mg·L-1IAA.After hardening,90% of plants were successfully established under greenhouse conditions.Histological observation revealed that shoot primordium originated from subepidermal cells of hypocotyl explants and directly developed into adventitious shoots without caHus formation. 相似文献
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Leaves of fine Populus tomentosa genotype TC152 were used as explants to establish cell suspension lines. The effects of plant growth regulators on callus induction and establishment of cell suspension lines were studied. The callus induction rate was the highest on a MS solid medium supplemented with 1.0 mg·L-1 2,4-D. A cell suspension line could be obtained by inoculating calli which were not subcultured into a MS liquid medium supplemented with 1.5 mg·L-1 2,4-D. The best subculture medium was MS + 0.8 mg... 相似文献
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Zhao Peng Dong Zhan-yuan Sun Hong-bin Zhao Ju-ying Wang Hua-fang College of Biological Sciences Biotechnology Beijing Forestry University Beijing P. R. China Ecology & Environment College Inner Mongolia Agricultural University Huhhot P. R. China Alashan Australian Affairs Office Bayanhaote P. R. China Institute of Forestry of Alashan Inner Mongolia Alashan P. R. China 《中国林学(英文版)》2006,8(4):10-14
The purpose of our study was to establish a regeneration system for micropropagation of Populus euphratica Olivier. On the basis of an analysis of plant leaf mineral nutrients, a special medium was proposed, called MP2. In optimizing media for in vitro plant cultures including MS, B5 and MP2 media we employed hormones, auxin IAA, cytokine benzyladenine (BAP) and gibberellic acid (GA) in our factorial experiments on media. Adventitious shoots were derived from cuttings of adult plants taken from Xingjiang, west China, on selected media with MP2 0.5 mg·L-1 BA 0.1 mg·L-1 NAA. The shoots were elongated on a medium with 0.25 mg·L-1 BAP, 0.1 mg·L-1 NAA and 2 mg·L-1 GA and were then rooted on a medium with 0.2-0.5 mg·L-1 IBA. All the media were incorporated with 30 g·L-1 sucrose and an adjusted pH at 6.3. 相似文献
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以钟花樱(Cerasus campanulata)的嫩枝作外植体进行组织培养技术研究。结果表明：适合不定芽诱导的培养基为 MS+6-BA 1.0 mg · L-1+NAA 0.1 mg · L-1+蔗糖30 g · L-1+卡拉胶6.5 g · L-1,诱导率为44.44%,适合增殖培养的培养基为 MS+6-BA 1.0 mg · L-1+NAA 0.05 mg · L-1+GA30.5 mg · L-1+蔗糖30 g · L-1+卡拉胶6.5 g · L-1,增殖倍数为3.70,适合生根培养的培养基为1/2 MS+IBA 1.5 mg · L-1+NAA 0.1 mg · L-1+蔗糖30 g · L-1+卡拉胶6.5 g · L-1+活性碳0.5 g · L-1,生根率为82.23%。 相似文献
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In vitro cultured shoots of three Chinese poplar species (Populus tomentosa, P. alba cv. Pyramidalis andP. euphratica) and an improved poplar (FS-51), as a control, were exposed to the stress of low osmotic potential, salinity and high pH,
and the shoot growth was measured.P. tomentosa andP. euphratica could grow even under the high osmotic stress (0.5m mannitol; −1.7 MPa).P. euphratica shoots which were previously cultured on the medium containing 0.7m mannitol (−2.4 MPa) recovered its growth after being transplanted to the medium without mannitol. The shoot growth ofP. euphratica was not so reduced even on the medium containing 100 mM NaCl. However, growth of the other poplars markedly reduced with
10 mM NaCl. Medium pH (pH 5.8–10) also did not affect the shoot growth ofP. euphratica. These poplars have different tolerance to the stresses in cultured shootin vitro. P. euphratica was extremely tolerant to the stresses which are possible to damage plants in dry land.
A part of this study was presented at the 104th Annual Meeting of the Japanese Forestry Society. (1993). 相似文献
9.
以毛白杨为转基因受体材料,利用根癌农杆菌介导法转化来源于益母草的阳离子抗菌肽基因LJAMP2。经卡那霉素筛选,共获得50株抗性植株。GUS组织染色和PCR检测显示有30株抗性植株呈阳性,初步证明外源目的基因已整合到毛白杨基因组中。RT-PCR证实抗菌肽基因LJAMP2在转基因植株中能大量表达。离体抗病性试验表明:转基因毛白杨细胞粗提液的抑菌能力明显强于非转基因植株。进一步将溃疡病菌接种在转基因和野生型毛白杨茎段上培养30天,转基因植株的病级指数均低于非转化植株。上述抗性试验结果表明:在毛白杨中超量表达益母草抗菌肽基因LJAMP2能显著提高其溃疡病抗病性。 相似文献
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以巨桉栽培无性系EG5无菌苗的叶片为外植体进行愈伤组织诱导与植株再生研究。结果表明:愈伤组织高效诱导和不定芽分化的最适培养基为改良MS+0.12 mg·L-1TDZ+0.25 mg·L-1NAA;硝酸铵对EG5叶片愈伤组织生长及植株分化的影响较大,最适宜质量浓度为0.412 5 g·L-1;最佳伸长培养基配方为改良MS+0.3 mg·L-16BA+0.05 mg·L-1IBA+0.3 mg·L-1IAA;暗培养10 d能促进不定芽分化,延缓愈伤组织老化和褐变速度。生根培养基为改良MS+0.4 mg·L-1NAA时生根率最高,为65.47%,移植成活率为90%以上。 相似文献
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农杆菌介导慈竹4CL基因遗传转化梁山慈竹 总被引：4，自引：0，他引：4
以梁山慈竹2种类型成熟胚的愈伤组织为材料,采用农杆菌遗传介导的方法,将已构建好的具有降低木质素含量的PBI121-4CL-RNAi表达载体导入愈伤组织,探讨愈伤组织预培养时间、菌液浓度、侵染时间、共培养时间和温度对遗传转化的影响。研究结果表明,淡黄色、颗粒状、疏松易碎的胚性愈伤组织是较好的遗传转化材料。以在愈伤组织培养基上预培养8天的淡黄色、颗粒状、疏松易碎的胚性愈伤组织为转化受体,在菌液浓度为OD600=0.05的EHA105中侵染20min后,在25℃、黑暗条件下共培养2天(共培养基表面加一层无菌滤纸),在含有卡那霉素为55mg.L-1的抗性筛选培养基上筛选30天,抗性愈伤组织率为90%,经PCR检测,慈竹4CL基因已导入梁山慈竹愈伤组织中。抗性愈伤组织在芽诱导培养基上诱导30天,可获得丛生芽,待丛生芽长至3～5cm后,在生根培养基上经过20～30天的诱导,可产生1～8条根,获得再生植株。经PCR检测,慈竹4CL基因已导入梁山慈竹再生植株中,获得了转基因植株,转化效率为9%。RT-PCR检测结果表明,转4CL基因的梁山慈竹愈伤组织和植株的内源4CL基因的表达受到抑制,且表达量比对照明显降低。 相似文献
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以感染丛枝病的豫杂一号泡桐幼苗为试验材料,通过不同浓度5-氮胞苷和利福平处理,研究了其幼苗形态和蛋白质的变化。结果表明,适宜浓度的利福平和5-氮胞苷共同使用可抑制泡桐丛枝病的发生。150μmol·L-15-氮胞苷+80 mg·L-1利福平可以抑制豫杂一号泡桐丛枝病症状。存在于健康豫杂一号幼苗中的蛋白质pI 6.8、m24ku而在病苗中观察不到。表明该蛋白质可能与豫杂一号泡桐丛枝病发生有一定关系。 相似文献
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以2 a托里桉超级苗不同株系的茎段为外植体进行组培快繁研究,结果表明：适合诱导的培养基为MS改良+6-BA 0.5mg·L-1+NAA 0.1 mg·L-1;适合6号株系增殖培养基为MS改良+6-BA 0.2 mg·L-1+NAA 0.1 mg·L-1,适合7号、10号株系的增殖培养基为MS改良+6-BA 0.3 mg·L-1+NAA 0.2 mg·L-1;适当缩短继代周期是解决桉苗顶稍枯萎、上部叶片脱落等症状简单、有效的方法;适合的生根培养基为1/2MS改良+ IBA 0.2 mg·L-1+ ABT 0.6 mg·L-1,生根苗移栽成活率可达90%。 相似文献
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Colt cherry (Prunus avium x pseudocerasus) callus cultures were derived from leaf protoplasts, protoplasts of root cell suspension cultures, or by direct culture of leaf and root tissues. Survival of calli cultured on basal proliferation medium containing 25, 50, 100 or 200 mN (millinormal) NaCl, Na(2)SO(4) or KCl, or iso-osmotic (with NaCl) concentrations of mannitol ranged from 1 to 15%. After six transfers on the same medium, surviving cell lines were subjected to three cycles of direct recurrent selection; i.e., in each cycle, they were cultured alternately on basal proliferation medium, and on basal proliferation medium supplemented with NaCl, KCl, Na(2)SO(4) or mannitol. Salt- or mannitol-tolerant cell lines selected in this way had smaller cells than unselected cell lines, and they grew more rapidly and had higher callus and cell survival rates than unselected cell lines when cultured in the presence of salt or mannitol. Cells lines selected for tolerance to one agent (sodium salt, potassium salt or mannitol) showed minimal tolerance to another agent. However, when plants were regenerated from salt- or mannitol-tolerant callus and new cultures derived from them, the new cultures showed tolerance to all of the salts and mannitol. Plant regeneration from the new cultures was not achieved under the conditions that led to the regeneration of the parent plants from callus. 相似文献
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以成年日本樱花茎段为外植体,系统研究了取材时间、冷藏时间、消毒方法、激素等诸多因素对茎段再生的影响,建立了日本樱花茎段丛生芽再生体系。研究结果表明,一般中午取材、4℃冷藏2 d、75%酒精浸泡3 min后用0.1%HgCl2浸泡7 min,可获得日本樱花无菌材料。将无菌材料接种至MS+NAA0.05 mg.L-1+6-BA2.0 mg.L-1的分化培养基上,腋芽明显萌动并伸长;在MS+6-BA4.0 mg.L-1+NAA0.3 mg.L-1+GA30.05 mg.L-1的增殖培养基上,腋芽不但能较快增殖形成大量丛生芽,而且形成的小芽能继续长大。待小芽长至3 cm~3.5 cm时,将其切割下来转移至1/2MS+NAA0.6 mg.L-1+IBA0.2 mg.L-1生根培养基中,最终获得日本樱花的完整植株。 相似文献
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Protoplasts were isolated from the leaves of sterile plants ofPopulus euphratica Oliv. by using 1% Cellulase “Onozuka” RS and 0.25% Pectolyase Y-23 in 0.6m of mannitol solution. Protoplasts were cultured in modified Murashige and Skoog's (MS) medium which contained no ammonium
ions but was supplemented with BAP (6-benzylaminopurine), 2,4-D (2,4- dichlorophenoxy-acetic acid), and 1% sucrose at the
cell density of 9×104/ml. Cell divisions occurred in every culture medium, especially in the medium containing 0.5 mg/l of BAP and 0.1 mg/l of
2,4-D, in which callus was successfully induced by successive culture through cell cluster formation. Shoots were regenerated
from the callus, and their growth was enhanced on 1/2 MS medium containing 0.8 mg/l of BAP. Finally, shoots were rooted and
plantlets were regenerated on 1/2 MS medium without a hormone.
A part of this paper was presented at the 106th Annual Meeting of the Jpn. For. Soc. (1995). 相似文献
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花曲柳体胚发生和植株再生 总被引：1，自引：0，他引：1
以花曲柳合子胚的单片子叶为外植体成功诱导出体胚并获得再生植株。未成熟合子胚的子叶在添加400mg·L-1水解酪蛋白、0.25mg·L-16-BA、1.5mg·L-1NAA、70g.L-1蔗糖和6g·L-1琼脂的MS1/2培养基上可以成功诱导产生体胚,诱导率达到34.7%,每个外植体上体胚数量为2～9个。成熟合子胚的子叶在添加0.25mg·L-16-BA、2mg·L-1NAA的MS1/2培养基(其他成分同上)上可以成功诱导产生体胚,诱导率为10.0%。体胚在MS1/2培养基上经过成熟培养后可以正常萌发,萌发率87.6%。萌发的体胚植株在MS1/2+0.01mg·L-1NAA培养基上生长较好,具备实生幼苗的外观特征。经炼苗后的体胚苗移植到栽培基质(草炭土:蛭石:珍珠岩体积比为5:4:1)中可以正常生长,成活率为75.0%。 相似文献
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利用农杆菌介导法获得转codA基因麻竹再生植株的研究 总被引：2，自引：0，他引：2
温度是影响植物生存和生长发育的基本环境因子之一。大多数植物对低温等都是高度敏感的,低温伤害现象尤为突出,几乎涉及所有的经济植物。因此,改善植物的抗低温冻害的胁迫能力,可以显著提高植物的生长范围、增加产量。codA基因可以增加植物对低温胁迫的耐受能力,而Rd29A是一种胁迫诱导特异表达启动子,胁迫条件可以快速诱导基因表达,也可以减少由于转基因过量表达带来的不利影响。研究以麻竹花药离体培养的愈伤组织为材料,采用农杆菌介导法,探讨了影响麻竹愈伤组织遗传转化效率的主要因子。结果表明,潮霉素的最佳筛选浓度是25 mg.L-1,预培养时间为3 d,侵染时间为20 min,共培养时间为3 d,乙酰丁香酮的浓度控制在100 mg.L-1时可以有效的提高遗传转化效率。在此基础上对获得的转基因植株进行分子检测,初步表明外源基因codA已经整合到麻竹基因组中。 相似文献
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报道细胞分裂素和硝酸铵对欧洲云杉不定芽伸长过程的影响.采用5年生的欧洲云杉1个无性系不定芽丛来研究玉米素和硝酸铵对不定芽生长的影响.试验3种诱导类型(玉米素浓度分别为6.84、13.68、20.52 μmol·L-1,激动素浓度0.69 μmol·L-1,培养4周),玉米素最高浓度处理诱导的伸长芽数量最多,在4个无生长调节剂继代培养过程中,加入了1个硝酸铵浓度逐渐增加的继代培养过程.经不含生长调节剂的低浓度的硝酸铵(2.5 mmol·L-1)培养基的继代培养后转入含有细胞分裂素的诱导培养基,下一个继代培养基补充的硝酸铵浓度分别为2.5、7.5、12.5、17.5 mmol·L-1,8周(2个继代培养)后转入含有硝酸铵浓度为2.5 mmol·L-1的培养基培养.硝酸铵浓度为7.5 mmol·L-1的处理可形成数量最多的不定芽,这对于采用不同浓度的玉米素进行诱导的处理结果是一致的,但随着硝酸铵的浓度增至12.5 mmol·L-1和17.5 mmol·L-1时则不定芽的数量减少.欧洲云杉的胚轴外植体经1次或多次诱导后,细胞分裂素(BA、激动素、玉米素、2iP)促进了不定芽的分化和发育,其中BA由于可诱导产生大量的不定芽而成为最有效的细胞分裂素.玉米素虽然诱导不定芽相对弱一些,却在BA多次诱导后可促进芽的伸长.这对获得具有形成不定芽能力并可扩繁的芽丛非常重要. 相似文献