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利用在Genebank上已报道的香石竹ACC氧化酶(ACO)基因的CDNA序列设计特异引物,以香石竹品种'MASTER'的eDNA为模板进行PCR扩增,克隆香石竹ACC氧化酶(AC0)基因.测序结果显示,克隆基因的序列与报道序列完全一致.将克隆的ACC氧化酶(AC0)基因分别连接到植物表达载体PBI121启动子CaMV 35S的上游及下游,构建香石竹ACC氧化酶(Aco)基因的正义表达栽体PBI121-ACO及反义表达载体PBI121-anti ACO.经PCR鉴定,基因已成功构建到表达裁体上.为香石竹抗衰老基因工程育种奠定了基础. 相似文献
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牡丹ACC氧化酶基因的克隆与反义载体的构建 总被引:1,自引:0,他引:1
根据报道的牡丹ACC氧化酶基因(DQ337251)cDNA序列,设计一对特异引物,以牡丹品种"洛阳红"基因组DNA为模板,用PCR扩增方法克隆出牡丹ACC氧化酶基因的部分片段,并将其连接到pMD18-T载体上进行测序.结果表明,克隆的序列全长为467 bp,其中包括一个长度为157 bp的序列,推测它可能是一个内含子,其它序列与已报道序列同源性为98.2%;用SacⅠ和XbaⅠ对重组质粒和载体pBI 121酶切、连接,构建牡丹ACC氧化酶基因的反义表达载体. 相似文献
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为探索番茄果实成熟与乙烯的关系,测定了转反义NR和ACC基因突变体番茄果实不同发育期乙烯释放量的变化,结果显示:(1)NR2、NR17和NR18号转反义NR基因果实各期的乙烯含量变化规律与普通果实相同。(2)转反义 NR基因番茄果实与转反义ACC基因番茄果实不同,前者果实能正常成熟、但成熟延迟,后者不能正常成熟。(3)在转反义NR基因果实发育各期中,粉红期乙烯释放量最高。(4)转反义NR基因果实成熟各期乙烯的释放量均明显低于普通果实。说明乙烯受体蛋白基因——NR基因的表达在被抑制后,也反过来抑制了番茄果实乙烯的释放。(5)转反义NR基因果实在常温下贮藏,贮藏期延长了43d。 相似文献
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应用反义RNA技术抑制甜瓜成熟过程中内源乙烯的合成,从而培育耐贮运品种是解决甜瓜延熟保鲜难题的可行新方法。根据GenBank中甜瓜、黄瓜ACC合成酶基因氨基酸保守序列设计引物,从成熟的薄皮甜瓜(齐甜1号)果肉组织中提取总RNA,经RT-PCR扩增得到约0.7kb的ACC合成酶cDNA片段,将其克隆到质粒载体pGEM-TEasy中,测序表明,该基因为777bp,编码258个氨基酸;从番茄(东农706)叶片组织中提取总DNA,经PCR扩增得到约2.2kb的E8基因片段,将其克隆到质粒载体pGEM-TEasy中,测序表明,该基因为2192bp;以pCAM2301为起始植物表达载体,pCAM-GT为中间载体,成功构建了果实特异启动子(E8)调控薄皮甜瓜ACC合成酶cDNA反义表达载体,采用冻融法将其转入根癌农杆菌LBA4404,得到了完整的Ti质粒表达载体系统。 相似文献
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香蕉ACC氧化酶基因(MAO3)的克隆及其表达特性分析 总被引:3,自引:0,他引:3
根据同源扩增得到香蕉(Musa acuminata) ACC氧化酶基因(MAO3) 的核心部分, 再通过3'和5'RACE扩增上、下游序列以及Genome-Walker的方法得到启动子部分, 共获得3 718 bp长度的序列。将所得结果进行聚类分析, 发现香蕉中ACC氧化酶的氨基酸序列非常保守, 各序列间的同一性高达99% ,与单子叶植物和双子叶植物中ACC氧化酶的氨基酸序列的同源性在66.7%~71.8%之间。组织原位杂交试验表明MAO3基因的表达具有组织特异性, 初步认为是在韧皮部筛管组织中特异表达。运用实时荧光定量PCR技术, 实时监测到了MAO3基因和香蕉乙烯受体基因ERS2受机械伤诱导的定量变化。 相似文献
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以香石竹(DianthuscaryophyllusL.)为材料,克隆得到1个ERF转录因子基因,命名为DcERF-1。氨基酸序列比对及系统进化树分析发现,DcERF-1与拟南芥AtERF-1的同源性最高,属于Ⅸ(B3)亚组。实时荧光定量PCR显示,DcERF-1表达量在花中最高,且在花瓣自然衰老和乙烯诱导的衰老过程中都呈现先上升后下降的趋势。DcERF-1定位于细胞核中。DcERF-1在香石竹花瓣中瞬时过量表达后,花瓣褪色速度明显延缓,离子渗透率明显降低;DcERF-1被瞬时沉默后,花瓣褪色速度显著加快,离子渗透率显著升高,衰老标志基因Dc SAG12表达量显著上调。酵母单杂交试验证明乙烯信号转导途径核心转录因子DcEIN3能直接结合DcERF-1的启动子。双荧光素酶瞬时表达试验证明DcERF-1能抑制乙烯生物合成途径中ACC氧化酶基因Dc ACO4的表达活性。综合结果表明Dc ERF-1负调控香石竹切花衰老。 相似文献
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脱落酸对桃果实成熟软化和乙烯生物合成的影响 总被引:1,自引:0,他引:1
以"白丽"桃为试材,分析了果实室温条件下贮藏过程中外源ABA(脱落酸)处理和NDGA(去甲二氢化愈创木酸,ABA生物合成抑制剂)处理后多聚半乳糖醛酸酶、果胶甲酯酶、ACC合成酶和ACC氧化酶含量的变化,探讨了在室温贮藏条件下ABA对果实成熟软化和乙烯生物合成的影响。结果表明:外源ABA处理可以提高多聚半乳糖醛酸酶和果胶甲酯酶的活性,加速果实硬度软化进程。外源ABA处理可以提高ACC合成酶和ACC氧化酶的活性,使果实乙烯释放量增加并且乙烯释放高峰提前出现。而NDGA处理则可以抑制多聚半乳糖醛酸酶的活性,果胶甲酯酶活性与清水相比虽然有降低,但抑制效果不显著。NDGA对果实硬度下降也有延缓作用。NDGA可以抑制ACC合成酶和ACC氧化酶的活性,从而降低乙烯释放量并且使乙烯释放高峰延时出现。 相似文献
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秋福红树莓叶片离体再生植株研究 总被引:1,自引:0,他引:1
以红树莓品种秋福(Rubus L.cv.Autumn Bliss)离体叶片为外植体,研究适合其再生植株的叶片部位、放置方式、植物生长调节剂以及暗培养时间等条件。结果表明,叶片外植体接种于MS+TDZ2.00mg/L+IAA0.10mg/L的培养基,暗培养2~3周转移至正常光照下培养效果最好,愈伤组织形成率、不定芽再生率和外植体不定芽数分别为100.00%、95.83%和(5.57±0.27)个。将再生芽接种于1/2MS+IBA0.10mg/L的培养基中,35d后生根率达100.00%。再生植株炼苗后移栽,30d时成活率达97.30%,成功地建立了红树莓叶片的离体再生体系。 相似文献
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Removal of the cut rose flower of ‘Forever Yours’ decreased water uptake by 20.4%, leaf removal 78.5%, and both flower and leaf removal 95.2%. Cut carnation ‘White Sim’ water uptake declined 27.1, 37,3 and 59.6% after flower, leaf, or both were removed, respectively. A high rate of water uptake (40.4%) continued after both carnation flower and leaf removal. 相似文献
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秋水仙素处理离体叶片获得皇家嘎拉苹果四倍体植株 总被引:30,自引:5,他引:25
采用皇家嘎拉苹果(Malus domestica Borkh.)离体新梢叶片作为外植体,研究了不同浓度的秋水仙素长时间处理(5 d)诱导四倍体的效率。结果表明,以不定芽再生培养基附加25 mg/L秋水仙素效果最好,最高在36.7%的叶片外植体上获得了四倍体植株。高浓度(75~200 mg/L)秋水仙素处理严重抑制组织的再生。采用流式细胞技术测定出诱变植株的细胞核DNA含量比对照高出一倍,确定了植株为四倍体。四倍体植株不仅在遗传上而且在外观形态上也明显区别于二倍体。四倍体植株已移栽于大田,并进行了嫁接育苗和大树高接。 相似文献
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Ehsan Ullah Khan Xing-Zheng Fu Jing Wang Qi-Jun FanXiao-San Huang Ge-Ning ZhangJie Shi Ji-Hong Liu 《Scientia Horticulturae》2009
In the current work attempts were made to investigate culture of leaf explants derived from in vitro seedlings of two sweet orange (Citrus sinensis (L.) Osbeck) cultivars, Bingtangcheng and Valencia. Effects of several factors, including culture medium, lighting condition, explant age and genotype on regeneration response were examined based on three parameters, percentage of explants producing shoots, mean number of shoots per explant and shoot forming capacity. Culture of the explants on shoot-inducing media (SIM) composed of MT salts supplemented with different growth regulators gave rise to disparate shoot regeneration, in which SIM1 (MT + 0.5 mg L−1 BA + 0.5 mg L−1 Kinetin + 0.1 mg L−1 NAA + 3% sucrose + 0.8% agar, pH 5.8) was shown to be the most effective medium for direct induction of shoots from leaf explants. Highly significant difference in the response of shoot bud regeneration was noted between the two cultivars, with Bingtangcheng being more responsive than Valencia. Culture of explants from fully developed leaves led to better shoot regeneration capacity in comparison to undeveloped ones. However, the two lighting conditions used herein did not cause significant difference in shoot regeneration. Phenotypic observation and randomly amplified polymorphic DNA (RAPD) analysis confirmed that all the regenerated plants from both genotypes were genetically identical to their donor plants, suggesting absence of detectable genetic variation in the regenerated plants. The data presented here demonstrated that direct initiation of plants from leaf explants has been successfully accomplished. To our knowledge, this is the first report on direct regeneration of shoots from leaf explants in Citrus, which will provide an alternative source for citrus genetic manipulation in the future. 相似文献
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红龙草叶片的组织培养及其植株再生 总被引:5,自引:0,他引:5
建立了红龙草叶片再生体系。叶片外植体在培养基MS + 6-BA 1.0 mg/L + 4-PU 1.0 mg/L +NAA 0.1 mg/L上形成浅绿色愈伤组织, 20 d后愈伤组织诱导率达100%。约45.31%的愈伤组织在添加6-BA 1.0 mg/L和NAA 0.4 mg/L的MS培养基上分化出紫红色的不定芽, 约6%的愈伤组织在该培养基上产生出细小叶片和绿色变异幼芽。所产生的紫红色不定芽在1/2MS +NAA 0.4 mg/L的培养基上可全部生根,长成完整植株。再生植株的移栽成活率达85%以上。 相似文献
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ABSTRACT: BACKGROUND: Quantification of leaf movement is an important tool for characterising the effects of environmental signals and the circadian clock on plant development. Analysis of leaf movement is currently restricted by the attachment of sensors to the plant or depended on visible light for time-lapse photography. The study of leaf growth movement rhythms in mature plants under biological relevant conditions, e.g. diurnal light and dark conditions, is therefore problematic. RESULTS: Here we present OSCILLATOR, an affordable system for the analysis of rhythmic leaf growth movement in mature plants. The system contains three modules: (1) Infrared time-lapse imaging of growing mature plants (2) measurement of projected distances between leaf tip and plant apex (leaf tip tracking growth-curves) and (3) extraction of phase, period and amplitude of leaf growth oscillations using wavelet analysis. A proof-of-principle is provided by characterising parameters of rhythmic leaf growth movement of different Arabidopsis thaliana accessions as well as of Petunia hybrida and Solanum lycopersicum plants under diurnal conditions. The amplitude of leaf oscillations correlated to published data on leaf angles, while amplitude and leaf length did not correlate, suggesting a distinct leaf growth profile for each accession. Arabidopsis mutant accession Landsberg erecta displayed a late phase (timing of peak oscillation) compared to other accessions and this trait appears unrelated to the ERECTA locus. CONCLUSIONS: OSCILLATOR is a low cost and easy to implement system that can accurately and reproducibly quantify rhythmic growth of mature plants for different species under diurnal light/dark cycling. 相似文献
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Vegetative shoot cuttings of 6 cultivars of carnation (Dianthus caryophyllus L.) were irradiated with 30 or 60 Gy X-rays. Subsequently, from both control and irradiated cuttings, explants were excised at 2 positions of the stem and cultured in vitro. The effects of X-ray dose, genotype and explant position on multiple shoot development of in vitro-cultured nodal stem explants, and on root formation of sub-cultured shoots, were examined. The in vitro propagation method resulted in the production of 2220 plantlets from 209 explants, 4 months after incubation. 相似文献