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利用转基因技术将多种抗病基因共同转入毛白杨中以提高其抗性,从而获得毛白杨抗病新品种是目前解决杨树真菌病害的主要研究方向之一。本研究通过根癌农杆菌介导的二次遗传转化,将来源于球孢白僵菌几丁质酶基因Bbchit1转入过量表达无色花色素还原酶基因LAR3转基因毛白杨中,实时定量PCR显示Bbchit1与LAR3均能有效表达,离体抗病试验显示Bbchit1+LAR3共表达转基因毛白杨细胞粗提液对杨树叶枯病菌具有明显抑制作用,进一步将叶枯病菌接种在转基因和非转基因毛白杨叶片上培养30天,转基因植株的感病面积均低于非转基因植株且Bbchit1+LAR3共表达转基因株系抗病效果更明显。上述抗病试验结果表明:LAR3和Bbchit1在杨树中共表达可提高其对叶枯病的抗性。 相似文献
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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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The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase (SPDS) genes derived from an apple by an Agrobacterium-mediatod method. Four transgenic clones were confirmed by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR. 相似文献
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Development of transgenic plants with tolerance to environmental stress is an important goal of plant biotechnology. Late-embryogenesis-abundant(LEA) proteins accumulate in seeds during late embryogenesis, where they protect cellular membranes and macromolecules against drought. In this work, we transferred the Tamarix androssowii LEA gene into hybrids of Populus davidiana×P. bolleana. We compared relative rates of height growth, chlorophyll fluorescence kinetic parameters, and leaf Na+ levels of six TaLEA-containing lines with non-transferred plants(NT), all grown under 0.8% NaCl stress condition. Survival percentages of transgenic lines were all higher than for NT controls after rehydration and the survival percentage of SL2 was five-fold higher than for NT controls. Seedling height increased 48.7% in SL2(from the onset of induced stress to the end of the growing season), 31% more than for the NT controls. Chlorophyll fluorescence kinetic parameters showed a marked increase in photosynthetic capacity in SL2 and SL5. Na+ levels in young leaves of transgenic lines were lower than in control NT leaves, but higher in yellow and withered leaves, indicating improved salt tolerance in transgenic lines. 相似文献
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Trehalose is a non-reducing disaccharide with high stability and strong water absorption properties that can improve the resistance of organisms to various abi-otic stresses.Trehalose-6-phosphate synthase (TPS) plays important roles in trehalose metabolism and signaling.In this study,the full-length cDNA of ThTPS was cloned from Tamarix hispida Willd.A phylogenetic tree includ-ing ThTPS and 11 AtTPS genes from Arabidopsis indicated that the ThTPS protein had a close evolutionary relationship with AtTPS7.However,the function of AtTPS7 has not been determined.To analyze the abiotic stress tolerance function of ThTPS,the expression of ThTPS in T.hispida under salt and drought stress and JA,ABA and GA3 hormone stimu-lation was monitored by qRT-PCR.The results show that ThTPS expression was clearly induced by all five of these treatments at one or more times,and salt stress caused par-ticularly strong induction of ThTPS in the roots of T.hispida.The ThTPS gene was transiently overexpressed in T.his-pida.Both physiological indexes and staining results showed that ThTPS gene overexpression increased salt and osmotic stress tolerance in T.hispida.Overall,the ThTPS gene can respond to abiotic stresses such as salt and drought,and its overexpression can significantly improve salt and osmotic tolerance.These findings establish a foundation to better understand the responses of TPS genes to abiotic stress in plants. 相似文献
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PtLFY, a LEAFY （LFY） gene, was cloned from Populus tomentosa （LM50） by PCR. Sequencing analysis indicated that PtLFY was 2629 bp long, composed of three exons and two introns and encoded 378 amino acids. The splice donor sites and the splice acceptor sites were in identical positions to the LFY and its homologues. The amino acid sequence inferred was 68%-99% homologous to those of LFY and its homologues by blast analysis in GenBank. The Southern blot analysis indicated that there was a single copy of the PtLFY gene in genomic DNA of male and female P. tomentosa （LM50 and 5082）. The pBI121-Ptalfy （reverse）-intron-Ptlfy-GUS-nos was constructed using RNA interference （RNAi） technique and verified by PCR and digestion identification and transformed into tobacco. Some transgenic tobacco plants were obtained by PCR and PCR-Southern identification. The growth was generally repressed in transgenic tobacco plants compared with wild-type ones and some phenotypic differences were observed. 相似文献
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经苗期对比试验,从全国１０省种源的１０４７株毛白杨优良基因资源中初选出３４个无性系进行中试,以比较速生的天然三倍体毛白杨雌株做对照。９年测定结果表明,鲁毛５０、５３１４、１４１４、５０１５和南×毛新五个无性系的材积增益高达３０％～５８．２％,且干形直,形态变异明显,易识别,适应性和抗逆性较强,有较高的推广利用价值。 相似文献
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Li Min Wang Hua-fang Yin Wei-lun He Si-jie Chen Shou-yi College of Biological Sciences Biotechnology Beijing Forestry University Beijing P. R. China Institute of Genetics Developmental Biology Chinese Academy of Sciences Beijing P. R. China 《中国林学(英文版)》2006,8(4):20-24
Transgenic lines were achieved by transforming the E. coli 1-phosphate mannitol dehydrogenase gene (mtl-D) into the Populus tomentosa Carr. genome. An Agrobacterium tumefaciens strain (AGL1), constructed by cloning mtl-D into the disarmed plasmid pBin438, was used to infect leaves of the clone YW2. The infected leaf discs were cultured on a medium containing 30 mg·L-1 kanamycin and 500 mg·L-1 cefotaxime. Transgenic plantlets regenerated from the infected leaves, rooted on the medium containing 30 mg·L-1 kanamycin. PCR and a Southern blotting test verified that the exogenous mtl-D gene had integrated into the transformation plants of the P. tomentosa genome. The mannitol content in control plant was 69μg·g-1 FW, and the mannitol contents of the transgenic lines T1 to T5 ranged between 103.7 and 289.5μg·g-1 FW. Of the shoots of the control plants 20% survived; on the medium containing 0.6% NaCl, 60% and 70% of two transgenic shoots survived on a medium containing 0.8% NaCl. 相似文献
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对转豇豆胰蛋白酶抑制剂基因的三倍体毛白杨杂种 [(毛新杨×毛白杨 )×毛白杨 ]的可溶性总蛋白和胰蛋白酶抑制剂蛋白的含量进行测定 .结果发现 ,与未转基因植株相比 ,所有供试转基因株系叶片中的总可溶性蛋白含量明显增加 ,但老叶的含量高于嫩叶 ,这表明转基因株系可溶性总蛋白含量增加可能是CpTI基因表达的结果或由于外源基因导入后引起杨树基因组中某些自身基因的表达所致 .转基因株系叶片中有较高含量CpTI,而对照叶片则检测不到CpTI,这进一步证实了CpTI基因已在转基因株系中得到稳定表达 .进一步比较发现 ,在供试的 5个无性系中 ,TG0 7、TG0 4和TG71在总蛋白含量和CpTI含量增加较为明显 .另外 ,聚丙烯酰胺凝胶电泳胶分析发现 ,与未转基因植株相比 ,在所有供试转基因株系叶片中均出现一条分子量为 11.3kD的清晰蛋白带 相似文献
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Verification of the resistance of a LEA gene from Tamarix expression in Saccharomyces cerevisiae to abiotic stresses 总被引：1，自引：0，他引：1
The role of late embryogenesis abundant （LEA） proteins in stress tolerance was examined by using a yeast expression system. LEA protein tolerance to the abotic stresses in plants involved in salt, drought and freezing stresses and additional tolerance to heat, NaHCO3 （salt-alkali） and ultraviolet radiation was also investigated. The transgenic yeast harboring the Tamarix LEA gene （DQ663481） was generated under the control of inducible GAL promoter （pYES2 vector）, yeast cells transformed with pYES2 empty vector were also generated as a control. Stress tolerance tests showed that LEA yeast transformants exhibited a higher survival rates than the control transformants under high temperature, NaHCO3, ultraviolet radiation, salt （NaCl）, drought and freezing, indicating that the LEA gene is tolerant to these abiotic stresses. These results suggest that the LEA gene is resistant to a wider repertoire of stresses and may play a common role in plant acclimation to the examined stress conditions. 相似文献
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To analyze the function of PtAP3, an APETALA3 (AP3) homologue gene isolated from Populus tomentosa Carr., the full length sequence (1 797 bp) and a fragment (870 bp) of PtAP3 were fused to a CaMV 35S promoter of pBI121 to generate the sense and antisense constructs of PtAP3. These constructs were transformed into tobacco by Agrobacterium infection of leaf disks and selection on kanamycin medium. Some sense and antisense transgenic tobacco plants were obtained by PCR and Southern blot analysis. Great phenotypic differences in transgenic tobacco plants were observed. Almost all of sense PtAP3 to transgenic tobaccos showed a higher growth rate than those of antisense transformants and a few developed pregnancy earlier than wild type seedlings and antisense transformants under the same conditions. 相似文献
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Wang Dong-mei An Xin-min Zhang Zhi-yi Key Laboratory of Genetics Breeding in Forest Trees Ornamental Plants Ministry of Education Beijing Forestry University Beijing P. R. China 《中国林学(英文版)》2005,(3)
To analyze the function of PtAP3, an APETALA3 (AP3) homologue gene isolated from Populus tomentosa Carr., the full length sequence (1 797 bp) and a fragment (870 bp) of PtAP3 were fused to a CaMV 35S promoter of pBI121 to generate the sense and antisense constructs of PtAP3. These constructs were transformed into tobacco by Agrobacterium infection of leaf disks and selection on kanamycin medium. Some sense and antisense transgenic tobacco plants were obtained by PCR and Southern blot analysis. Great phenotypic differences in transgenic tobacco plants were observed. Almost all of sense PtAP3 to transgenic tobaccos showed a higher growth rate than those of antisense transformants and a few developed pregnancy earlier than wild type seedlings and antisense transformants under the same conditions. 相似文献
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毛白杨抗寒无性系选育 总被引：4，自引：3，他引：1
利用全国毛白杨优树基因资源进行抗寒无性系选育,国内尚属首次。经８年系统研究,选育出１７个抗寒无性系,其中Ｌｘ、１１７１等７个无性系的材积超过小黑杨２３％以上,病虫害少,树干通直,无冻害。苗期复测和区域中试表明,这些抗寒无性系在北纬４０°左右,年均气温６℃以上,年平均极端最低温－２７．８℃和极值－３２．４℃的生态条件下,能正常发育生长并保持毛白杨特有的速生性,适宜在北方平川寒冷地区推广。而目前推广的三倍体毛白杨抗寒性差,冻害严重,只宜在热量条件较好的晋南推广。 相似文献
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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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表达豇豆胰蛋白酶抑制剂转基因杨树的虫试试验 总被引：2，自引：0，他引：2
将转豇豆胰蛋白酶抑制剂基因的三倍体毛白杨回交杂种 [(毛新杨×毛白杨 )×毛白杨 ]]于春季萌发前截干 .用萌条上的树叶对 3种主要杨树害虫 :天幕毛虫 (Malacosomadisstria)、舞毒蛾 (Lymant riadisparL .)和杨雪毒蛾 (StilpnotiacandidaStaudinger)进行离体虫试试验 ,年底测量各转基因株系萌条树高和地径 ,以衡量其生长性状 .结果发现 ,与对照相比 ,转基因株系的树高和地径没有显著降低 ,说明外源基因的导入没有影响转基因杨树的速生特性 ,相反 ,TG0 4的树高和地径较对照有明显的提高 ,这可能是外源导入后促进转基因杨树生长的结果 ;转基因植株均能显著提高害虫的死亡率 ,降低幼虫的排粪量、蛹重、体重增加量和成虫的产卵量 ;其中 ,TG0 7、TG0 8和TG713个转基因株系的抗虫特性较为突出 ,表明CpTI基因在它们的树体内表达较为活跃和稳定 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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Paper mulberry (Broussonetia papyrifera L. Vent) is well known for its bark fibers, which are used for making paper, cloth, rope, etc. It was found that, in addition to its well-documented role in the enhancement of plant salt tolerance, overexpression of the Na+/H+ antiporter (AtNHX5) gene in paper mulberry plants showed high drought tolerance. After exposure to water deficiency and salt stress, the wild-type (WT) plants all died, while the AtNHX5-overexpressing plants remained alive under high salt stress, and had a higher survival rate (>66%) under drought stress. Measurements of ion levels indicated that Na+ and K+ contents were all higher in AtNHX5-overexpressing leaves than in WT leaves in high saline conditions. The AtNHX5 plants had higher leaf water content and leaf chlorophyll contents, accumulated more proline and soluble sugars, and had less membrane damage than the WT plants under water deficiency and high saline conditions. Taken together, the results indicate that the AtNHX5 gene could enhance the tolerance of paper mulberry plants to multiple environmental stresses by promoting the accumulation of more effective osmolytes (ions, soluble sugars, proline) to counter the osmotic stress caused by abiotic factors. 相似文献