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西瓜和黄瓜乙烯受体ETR1基因片段的克隆与序列比较分析 总被引：2，自引：0，他引：2
乙烯受体基因ETR1是乙烯信号转导过程中的关键调控基因.研究根据ETR1基因的保守序列设计引物,以西瓜(Citrullus lanatus (Thunb.) Matsum & Nadai var.lanatus)和黄瓜(Cucumis sativus L.)的基因组DNA为模板进行PCR扩增,获得序列长度分别为1 633 bp和1 491 bp的基因片段CLETR1和CSETR1.序列分析表明,CLETR1和CSETR1与Genebank中收录的多条ETR1基因的核苷酸序列同源性在80%～98%,氨基酸序列同源性在75%～98%.西瓜和黄瓜ETR1基因片段的编码序列存在明显的单核苷酸变异,共23个核苷酸位点存在SNPs(Single nucleotide polymorphisms),其中5个SNPs导致4个编码氨基酸的改变. 相似文献
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从上西早生(Uenishiwase)柿叶片中,提取了基因组DNA。以基因组DNA为模板,经PCR扩增,克隆到一个549 bp的ETR5的DNA片段。序列分析表明,上西早生柿基因与洋梨ETR5的核苷酸同源性为79.0%,氨基酸同源性为81.0%;并且含有其他植物ETR5的保守氨基酸区和不变氨基酸残基,表明克隆到的序列是上西早生柿ETR5的基因片段。设计了2对带限制性内切酶位点的特异性引物,以测序质粒为模板,PCR扩增到2个ETR5-Uenishiwase片段。2个片段经单酶切消化后连接成反向互补的大片断。连接片断经双酶切消化后,连接到植物表达载体pBI221上,构建成Uenishiwase-ETR5基因的植物表达载体。 相似文献
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克隆与肥城桃成熟有关的乙烯受体ETR1基因片段,优化ETR1的表达条件,获得ETR1重组蛋白,为进一步研究ETR1功能并改善肥城桃贮运性能奠定基础。以肥城桃成熟果实总RNA反转录cDNA为模板,经PCR扩增得到特异片段,将该片段连接到质粒,经双酶切后,连接至质粒并转染至表达载体,IPTG诱导,优化表达条件。并以cD-NA为模板,进行实时荧光定量PCR,测定外源NO对ETR1相对表达量的影响。序列分析结果表明,该序列与Gen-Bank中的AF124527的cDNA序列同源性为99%,氨基酸序列同源性为99%。优化蛋白表达条件为:IPTG最佳浓度为0.5 mmol/L,最适温度为30℃,最适诱导时间为8 h。经优化后,ETR1重组蛋白成功表达,为TETR1的重组生产和对ETR1功能的体外研究提供了条件。此外,外源NO对体外表达的ETR1基因有抑制作用。 相似文献
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用莲雾的叶片提取基因组DNA作为模板,根据乙烯受体基因的保守序列设计引物,进行PCR扩增,得到长度约1.5kb的目的片段,将其克隆到pGEM T-easy载体转化大肠杆菌,提取质粒,进行限制性内切酶图谱分析及序列测定.结果表明,该目的片段全长为1400bp,与栽培稻(Oryza satzva)的osers基因组DNA可比对序列的一致性为98.68%.在所应用的12个内切酶中,该片段内部没有切点,根据序列特征和与其他乙烯受体基因序列比较,推测该片段有2个内含子和3个外显子片段组成,外显子总长为972 bp,编码324个氨基酸,其中第197～324氨基酸残基与组氨酸蛋白激酶的磷酸基团接受域有同源性. 相似文献
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本研究用自行设计的乙烯受体(ERS)保守引物从海南疣粒野生稻基因组DNA中克隆获得长度为1454bp的乙烯受体基因片段，推测该片段由2个内含子和3个外显子组成，外显子总长为974bp，编码324个氨基酸。与栽培稻(O.sativa)乙烯受体基因基因组DNA(os DNA)可比对序列的相似性达81．84％，推导出其mRNA与栽培稻的乙烯受体基因cDNA(OsERS)相应片段的同源性高达95．59％，而与栽培稻(Oryza sativa subsp Indica)ERS2相应片段的同源性为71．05％。 相似文献
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甘薯S-腺苷甲硫氨酸合成酶基因克隆与表达 总被引：6，自引：0，他引：6
从实验室建立的甘薯冷胁迫抑制消减文库中分离出一cDNA片段,经NCBI比对后发现该片段与其它植物的S-腺苷甲硫氨酸合成酶基因具有90%的同源性,根据相关物种S-腺苷甲硫氨酸合成酶基因cDNA序列设计引物,以甘薯总RNA为模板,经RT-PCR扩增首次获得全长1182bp的甘薯SAMS完全编码区,NCBI比对分析结果表明：该片段与不同种属植物sams基因的编码区序列的核苷酸相似性达85.48%, 所推导的氨基酸序列相似性达93.6%。根据核苷酸序列和氨基酸序列分别构建了SAMS系统进化树,从分子水平阐明了植物种属间的亲缘进化关系,为其种质资源利用提供理论依据。利用该序列与原核表达载体pET32a连接构建融合表达载体pET-SAMS,酶切鉴定后转入大肠杆菌BL21,在不同温度下诱导3h后均表达出一63KDa大小融合蛋白。 相似文献
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为揭示黄瓜绿斑驳花叶病毒的分子流行学特征,通过RT-PCR和cDNA克隆技术获得并分析了从北京甜瓜和山东西瓜上分离的2个黄瓜绿斑驳花叶病毒(CGMMV)分离物CGMMV-Beijing和CGMMV-Shandong的全序列.结果表明CGMMV-Beijing和CGMMV-Shandong的基因组分别由6423和6427个核苷酸组成,包括5'及3'端非编码区和4个开放性的阅读框,分别编码129kDa和186kDa复制相关蛋白、29kDa的移动蛋白及17.4kDa的外壳蛋白.寄主范围测定显示在供试的6科11种植物中CGMMV-Beijing和CGMMV-Shandong均可侵染葫芦科的葫芦、南瓜和西瓜以及茄科的本氏烟和藜科的苋色藜,不侵染供试的其它植物.CGMMV-Beijing和CGMMV-Shandong基因组的核苷酸同源性为99.5%,186kDa蛋白的核酸和氨基酸同源性分别为99.7%和99.8%,移动蛋白的核酸和氨基酸同源性分别为99.3%和98.1%,外壳蛋白的核苷酸同源性为100%.多序列比对分析结果显示,CGMMV的各分离物之间的分子差异并不明显,核苷酸同源性均在98%以上.将CGMMV-Beijing和CGMMV-Shandong与侵染葫芦科植物的烟草花叶病毒属其它成员比较,结果显示其亲缘关系较远,基因组的核苷酸同源性介于56.7%～59.6%,外壳蛋白的氨基酸同源性在44.7%～54.0%之间.综合分析显示,CGMMV分离物之间的差异并不明显,可能具有共同的侵染来源. 相似文献
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在先前的研究中通过抑制缩减杂交获得了一个在巴西橡胶幼态无性系与老态无性系胶乳中差异表达的片段(HbSSHl2),BlastX分析表明,由该片段编码的氨基酸与麻疯树(Jatropha curcas L.)翻译控制肿瘤蛋白(translationally controlled tumor protein,TCTP)的同源性达到92%.本研究根据HbSSHl2的序列信息设计引物,通过3L-RACE的方法获得了一个长832bp的cDNA(命名为HbTCVP),序列分析表明HbTCTP有507 bp的阅读框,68 bp的5'-UTR和255 bp的3'UTR,编码168个氨基酸,该氨基酸序列与麻疯树、油棕、花生、南瓜、番茄和拟南芥的TCTP同源性分别达到93.45%、90.48%、89.29%、85.12%、82.74%和79.76%.RT-PCR表明,HbTCTP在巴两橡胶叶片、胶乳及树皮中都有表达,乙烯利处理可诱导HbTCTP的表达,割胶抑制HbTCTP的表达,表明HbTCTP可能参与伤信号或乙烯信号传导.HbTCTP的表达分析有助于解析橡胶树幼态无性系高产的分子机制. 相似文献
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Ana Maria Borda David G. Clark Donald J. Huber Bruce A. Welt Terril A. Nell 《Postharvest Biology and Technology》2011,59(3):245-252
The relationship between fragrance and vase life and the role of ethylene on volatile emission in cut rose flowers was investigated. No relationship was observed between the amounts of volatile compounds emitted and vase life when fragrant and non-fragrant rose cultivars were compared. Neither ethylene production nor respiration rate of flowers was directly related with vase life. Volatile production during vase life was differential and independent among volatiles originating from different biosynthetic groups. Ethylene did not play a role in the regulation of volatile emission in rose flowers. Endogenous ethylene production was very low in most of the cultivars and did not show autocatalytic production trends. Volatile emission patterns during vase life did not parallel endogenous ethylene production. Exogenous ethylene exposure had differential effects among all cultivars, regardless of the fragrance of the flower. Fragrant cultivar ‘Osiana’ was highly sensitive to exogenous ethylene, with petals abscising within 24 h of ethylene (1 μL L?1) exposure while other fragrant cultivars ‘Erin’ and ‘Lovely Dream’ had low ethylene sensitivity. Volatile production was unaffected by exogenous ethylene. The results of this study indicate that volatile emission in cut roses is not regulated by endogenous or exogenous ethylene and occurs independently of petal senescence and/or abscission. These results provide a better understanding of the complexity of volatile emission in rose flowers. 相似文献
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Ethylene-mediated premature floral senescence and petal or flower abscission affect postharvest longevity of several species used as cut flowers. Exposure to exogenous or endogenously produced ethylene can be controlled in several ways. These include the use of ethylene biosynthesis inhibitors or ethylene action inhibitors, and ethylene removal technologies. In addition, genetic modification can be very effective in controlling ethylene synthesis and perception. We review here the potential for applications of nanotechnology to control ethylene levels and postharvest management in the flower industry. Already, nanosponges have been shown to enhance efficacy of the ethylene inhibitor 1-MCP in several flower species. In carnation, 1-MCP included in nanosponges also allowed better control of Botrytis cinerea damage. However other applications are also considered based on successes in the use of this technology to increase agricultural production and decrease postharvest waste. Nano-metal based sensors could be used for detection of ethylene in the store and to label the product along the distribution chain. Furthermore, nanocomposites could be included as scavengers for ethylene removal in active packaging, and nanocatalysts could promote ethylene catalytic degradation in the warehouse. Nanoparticles could also be introduced into a new generation of packaging to control effects of gases and UV, and increase strength, quality and packaging appearance. This review highlights recent results on the use of nanotechnology sensu lato and potential application for cut flower vase life improvement, focusing on ethylene control strategies. 相似文献
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玉米细胞质雄性不育性与乙烯的关系 总被引：13，自引：0，他引：13
研究了玉米Ｍｏ１７核背景三种雄性不育细胞质（Ｔ、Ｃ、Ｓ）系及正常可育细胞质（Ｎ）系小孢子发育不同阶段花药组织的乙烯释放量，发现不育系小孢子败育前的一、二个时期内花药组织乙烯释放量显著增加。同时设计的乙烯利及其合成抑制剂外源喷施实验也表明了其对雄性育性的逆转效应。讨论了植物激素对玉米细胞质雄性不育性的调控作用 相似文献
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Postharvest quality of cut lily flowers 总被引：1，自引：0，他引：1
This paper reviews the literature on the postharvest quality of cut lily inflorescences and highlights the need for more information. The main symptoms that limit the length of vase life are abscission of floral buds, lack of flower opening, tepal wilting, and leaf yellowing. Floral bud abscission is regulated by ethylene and can be prevented by treatments with inhibitors of the ethylene receptor. Lack of bud opening is also mimicked by ethylene treatment. It is alleviated by treatment with sugars. These might reduce ethylene effects or act as a source of energy. Depending on the cultivar, tepal wilting is not or is only slightly affected by ethylene. The time to tepal wilting is positively correlated with the levels of endogenous sugars. Leaf yellowing is apparently not affected by ethylene, and is aggravated by the inclusion of sugars in the vase solution. A treatment with hormones, in particular GA4+7 and benzyladenine, can prevent or alleviate the negative effects of sugars on leaf quality.A relatively short period of cold storage often drastically increases the number of floral buds that fail to open. Cold storage also hastens tepal wilting, induces or increases leaf yellowing, and promotes bud abscission. Several lily hybrids, therefore, seem chilling-sensitive. Some of the negative effects of cold storage can be alleviated by sugars, and others by GA4+7, with or without benzyladenine.The lack of ethylene sensitivity of tepal senescence, in several, but not all, cultivars, implies that pollination will also not affect senescence. It would be interesting to test this hypothesis in a few cultivars. It is also not clear if ethylene (or pollination) has an effect on tepal abscission in cut lilies. More data are also needed with regard to chilling injury. Pulsing with sugars might, for example, reduce the degree of injury. 相似文献
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农作物乙烯合成和信号转导途径及其对抗病反应的调控 总被引：1，自引：0，他引：1
乙烯作为植物激素在生长、发育、抗逆过程中发挥了其独特的调节作用。本文在分子生物学水平上概述了植物中乙烯合成途径和信号转导途径的机制。乙烯的合成由甲硫氨酸开始,经过重要的中间代谢产物ACC的氧化裂解形成乙烯,其中ACC合成酶催化的反应为限速反应,为调控乙烯合成的重要环节。乙烯信号的转导由内质网上乙烯受体识别乙烯开始,在胞质中经一条保守的途径,由EIN3将转录信号传递至细胞核中,最后以ERF类转录因子激活或抑制相关基因的表达。ERF转录因子参与防卫反应的诱导和寄主对病原菌不亲和互作的建立,受其调控的防卫基因被诱导表达后在随后防卫应答过程中发挥了不同的作用。 相似文献
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Raspberries are fruit with high metabolism that makes them very perishable, impairing their storage and shelf-life. Chitosan coatings have the potential to improve their postharvest life by reducing water loss, respiration rate and decay incidence. The purpose of this work was to study the effect of different concentrations of chitosan, applied pre- or postharvest, on the retention of quality attributes of fresh raspberries. The chitosan concentrations tested were 0 (control), 0.5, 1.0 or 2.0%. The postharvest treatment was applied immediately after harvest, dipping the fruit in the solutions for 5 min. The pre-harvest treatment was done with one hand-spray application per week for three weeks, starting when the fruit were just turning pink. In both experiments the fruit were stored at 0 °C and 90% RH. Pre- or postharvest use of chitosan at 1 or 2% was effective in maintaining titratable acidity and retarding respiration and ethylene production, weight loss and decay incidence. Application by both means resulted in the highest chitosan concentrations accelerating a reduction of ascorbic acid contents. Firmness was maintained only when the fruit were treated pre-harvest at 2%. Thus, application of chitosan at 1 or 2% postharvest and 2% pre-harvest was able to retain key raspberry quality attributes for 15 and 12 days, respectively. 相似文献
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植物乙烯受体及转基因育种研究进展 总被引：8，自引：0，他引：8
在对模式植物拟南芥的遗传学和分子生物学的深入研究中,获得了乙烯应答过程中大量的突变体,分离了编码乙烯受体的基因,其编码产物的结构和功能也已得到鉴定,一些乙烯受体基因已用于转基因植物的研究。本文对近几年已确认的乙烯受体基因突变体,对乙烯受体基因的遗传途径、表达模式及其编码产物的结构、功能及其相互关系做了综述。探讨了利用乙烯受体基因进行转基因植物研究的可行性。 相似文献