共查询到18条相似文献，搜索用时 437 毫秒
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采用cDNA-AFLP技术分离了一个与番木瓜果实成熟衰老相关的差异片段,经生物信息学分析证实该片段属于半胱氨酸蛋白酶（Cysteine Protease,CP）基因。将差异片段序列在NCBI中的番木瓜基因组序列中搜索,获得了半胱氨酸蛋白酶基因DNA序列。设计开放型阅读框上、下游引物,以木瓜果肉RNA逆转录的cDNA为模板扩增该基因全长cDNA序列,命名为CpCP（登录号：JN689334）。该基因属于C1肽酶家族中的C1A亚家族,编码471个氨基酸,含有家族特有的催化三联体：Cys166-His302-Asn322。Real-time PCR结果显示CpCP在果实中大量表达,是根、茎、叶、花中表达量的几十倍;CpCP受乙烯处理诱导,受1-MCP处理抑制,表达模式与番木瓜果实成熟衰老进程一致,说明CpCP参与了番木瓜果实成熟衰老进程。 相似文献
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《果树学报》2010,(6)
以番木瓜果肉为试材,提取总RNA,反转录为双链cDNA。采用cDNA末端快速扩增方法(RACE),获得了番木瓜果肉Actin基因的全长cDNA序列,将其命名为CpActin,Genbank登录号为FJ696416。CpActin全长cDNA为1533bp,含有一个1134bp的开放阅读框,编码377个氨基酸,与毛白杨、葡萄、水稻、拟南芥的Actin同源性分别为98.94%、98.67%、96.29%、94.69%。利用邻接法构建了部分高等植物、哺乳动物和真菌的Actin系统进化树,结果表明番木瓜与葡萄的Actin进化距离最小。采用半定量RT-PCR技术分析CpActin基因在不同成熟度番木瓜果实中的表达情况,发现该基因的表达水平没有明显差异,表明CpActin基因可以作为内参,进行番木瓜其他果实基因差异表达的研究。 相似文献
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《果树学报》2019,(11)
【目的】番木瓜是典型的呼吸跃变型果实,外源乙烯处理使番木瓜呼吸跃变提前,促进果实成熟。分离番木瓜果实成熟相关miRNA,为深入了解呼吸跃变型果实的成熟分子机制奠定基础。【方法】利用高通量测序技术对乙烯(ETH)、1-MCP和清水对照(CG)处理的番木瓜果实进行miRNA和转录组高通量测序,然后对测序获得数据进行生物信息学分析,进行miRNA鉴定和靶基因预测,并与转录组测序结果进行关联分析。【结果】乙烯、1-MCP和对照处理分别获得10 734 196、16 486 803和16 067 290条纯净序列,共鉴定出523个miRNA。其中,已知miRNA个数为1-MCP(303)、CG(214)和ETH(239),新miRNA个数为1-MCP(184)、CG(188)和ETH(114)。与对照相比,在乙烯处理中上调和下调表达的miRNA分别是123和72条。靶基因预测共获得5 053个靶基因,KEGG功能富集分析显示它们参与了戊糖、葡萄糖醛酸转换、淀粉和蔗糖代谢、卟啉和叶绿素代谢、类胡萝卜素合成等代谢途径。筛选出的番木瓜果实成熟衰老相关候选miRNA,包含4个果实软化调控相关miRNA(miR167-y、miR4993-x、miR3946-x和miR5059-x)、3个果实颜色调控相关miRNA(miR4993-x、miR815-y和miR7810-x)、3个激素调控相关miRNA(miR4993-x、miR8004-x和miR9722-x)和4个转录因子调控相关miRNA(miR5641-y、miR9722-x、miR838-y和miR319-y)。【结论】筛选的番木瓜果实成熟衰老相关miRNA为今后果实成熟衰老调控网络研究提供了可能的线索。 相似文献
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【目的】克隆一个与番木瓜果实软化相关的扩展蛋白基因,分析其功能,明确其在不同组织器官和不同成熟时期果实的表达模式。【方法】基于对番木瓜果实转录组学的研究,筛选并克隆了一个与果实软化相关的扩展蛋白基因Cp EXPA2。采用同源序列对比和系统进化树分析对该基因进行序列分析;运用生物信息学的方法对该基因编码的蛋白进行结构分析;使用RT-q PCR方法分析Cp EXPA2基因在不同组织器官、不同成熟时期果实中的表达量;采用GY-4硬度计测定不同成熟时期番木瓜果实的硬度。【结果】Cp EXPA2基因的开放阅读框(ORF)为780 bp(Gene Bank登录号为MF662209),编码259个氨基酸。该基因编码的氨基酸序列具有典型的扩展蛋白保守结构:N端富含8个半胱氨酸、C端富含4个色氨酸和中间1个组氨酸功能域(His-Phe-Asp,HFD)。同源序列比对分析发现该扩展蛋白与山木瓜(ABF48653.1)、番茄(AAC64201.1)、草莓(AAF21101.1)、拟南芥(AAB38073.1)等扩展蛋白氨基酸序列有较高的同源性,分别为66.15%、64.86%、66.80%、65.00%。进化树分析表明,Cp EXPA2蛋白与拟南芥At EXPA6(U30480)、At EXPA16(NM_115407)关系最近。Cp EXPA2在不同组织器官中的表达量依次为成熟果实叶花茎根;在不同发育时期的果实中,Cp EXPA2在果皮的表达量相对高于果肉,且随着果实成熟软化程度提高,其表达量也相应提高。番木瓜果实的硬度随着果实的成熟呈逐渐下降趋势,在破色期开始硬度急剧下降,果实迅速软化。【结论】Cp EXPA2基因的扩展蛋白家族高度保守,且该基因编码的蛋白与山木瓜、番茄、草莓、拟南芥等扩展蛋白的氨基酸序列有较高的同源性。系统进化树分析发现番木瓜Cp EXPA2与拟南芥At EXPA6和At EXPA16亲缘关系较近。Cp EXPA2受乙烯诱导,且与果实发育进程有关,因此推测该基因可能在番木瓜果实成熟软化过程中发挥着重要的作用。 相似文献
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《果树学报》2021,(2)
【目的】明确甜樱桃PavSS基因家族在果实成熟过程中的功能。【方法】利用qRT-PCR技术分析了7个蔗糖合成酶基因PavSS1～7在果实发育过程中的表达情况,并分析了蔗糖对PavSS家族基因表达的影响,利用甜樱桃果实的VIGS技术分析了PavSS1与PavSS6基因影响果实成熟的功能。【结果】PavSS1和PavSS2基因在甜樱桃果实发育和成熟过程中均表达,且PavSS1基因表达量是PavSS2基因的10倍以上;PavSS1和PavSS2在果实发育前期表达量逐渐升高,后期表达量逐渐降低。PavSS3和PavSS5基因在整个果实发育期均低表达,且表达量差异不显著。PavSS4和PavSS7基因在果实发育前期表达量高,后期表达量低。PavSS6基因在果实发育初期低表达,随后表达量逐渐升高,并维持较高水平。外施蔗糖显著下调了PavSS1和PavSS6基因的表达,而PavSS2、PavSS3、PavSS4、PavSS5和PavSS7基因的表达变化不显著。沉默甜樱桃果实PavSS1基因抑制了甜樱桃果实成熟,包括果实硬度、ABA含量、花色苷含量和可溶性糖含量发生显著变化以及成熟相关基因的表达量显著下调;而沉默PavSS6基因的甜樱桃果实与空载对照相比,没有显著变化。【结论】PavSS1基因可能是控制甜樱桃果实蔗糖代谢的关键基因,参与了甜樱桃果实成熟过程的调控。 相似文献
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香蕉泛素结合酶基因与果实成熟关系的研究 总被引：1，自引：1，他引：0
根据香蕉果实采后早期成熟的抑制缩减杂交文库获得的香蕉泛素结合酶基因片段,从香蕉（Musa acuminate L. AAA group‘Brazilian’）果实中克隆了泛素结合酶基因的cDNA全长,命名为MaUCE1。该cDNA全长890 bp,编码152个氨基酸。BlastX分析表明,该基因所推导的氨基酸序列与烟草（BAB40310）、小麦（AAA34310）、拟南芥（L19351）和马铃薯（ABA46759）有较高的一致性（95.39%、95.39%、92.11%和90.79%）,并且结构域分析发现该序列具有泛素结合酶E2的酶活性中心部位和一个与其他真核生物泛素E2酶相同,在泛素E2硫酯键形成中具有催化活性,并在进化上高度保守的半胱氨酸残基。该基因在香蕉根、茎、叶、花、果实中均有表达,但在花和果实中的表达量较高,并且在果实成熟后期表达量升高,与淀粉磷酸化酶活性变化一致,与淀粉含量的变化呈负相关,暗示该基因可能参与香蕉果实成熟过程中的分解代谢,而与果实成熟启动无关。 相似文献
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‘短柄樱桃’花芽休眠解除过程中差异表达基因的研究 总被引：1，自引：0，他引：1
以中国樱桃的优良品种‘短柄樱桃’（Prunus pseudocerasus Lindl.‘Duanbing’）的花芽为材料,采用mRNA 差异显示技术,分析休眠解除的过程中相关差异表达基因。共克隆获得79 个差异cDNA片段;通过半定量RT-PCR 验证,确定18 个阳性差异cDNA 片段为休眠解除相关候选基因;休眠解除过程中上调的基因片段11 个,下调的7 个。测序及同源性比对结果显示：差异表达基因主要参与糖代谢、信号转导、跨膜转运及氧化还原等反应过程。这些基因在‘短柄樱桃’休眠解除阶段可能起到直接或间接的作用。 相似文献
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设计植物EXP扩展蛋白简并引物,以砂梨果实cDNA为模板,克隆得到EXP基因cDNA片段,该片段长465 bp。根据该片段序列,分别设计2条5’和3’末端扩增的特异引物,利用RACE技术,获得了该片段的5’端和3’端序列。用DNAMAN5.22软件对3个序列进行拼接和分析,获得了该基因的cDNA全长,命名为Pyp-EXP。该cDNA全长为1 395 bp,5’端起始密码子ATG始于72 bp,3’端终止子TAG止于830 bp,Ploy+(A)从1 363～1 395 bp。该基因已在GenBank基因数据库注册,注册号为EF602031。Pyp-EXP核苷酸序列有一个759 bp完整的开放阅读框,编码区与西洋梨、苹果、湖北海棠、桃的同源性分别为96%,96%,94%和86%;该cDNA推导编码252个氨基酸,含有1个组氨酸(His_Phe_Asp,HFD)功能域,与西洋梨、苹果、湖北海棠、桃中相应序列同源性分别为98%,97%,95%和93%。该基因的克隆为研究扩展蛋白的时空表达及其在果实发育和成熟过程中的作用奠定了基础。 相似文献
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杨梅酸性转化酶基因cDNA分离及表达分析 总被引：1，自引：0，他引：1
杨梅果实富含蔗糖,酸性转化酶是蔗糖代谢关键酶,根据植物酸性转化酶基因保守区序列设计引物,提取杨梅叶片RNA,逆转录获得cDNA,以此为模板通过PCR技术扩增到长度为516bp的基因片段,克隆入pMD18-T载体中,命名为MrIVR1(GenBank:DQ339699)。测序及同源性检索表明,该基因推导氨基酸序列与君子兰、葡萄、草莓、胡萝卜等酸性转化酶基因氨基酸序列同源性为60%～69%。运用ClustalX软件对植物转化酶基因进行了系统树分析,结果显示,MrIVR1编码的蛋白质属于细胞壁酸性转化酶。半定量RT-PCR表达分析显示,MrIVR1基因在杨梅果实发育早期表达量最高,随着果实的发育表达量下降,在成熟果实中表达水平较低。 相似文献
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J. J. Geng Y. H. Shen F. Y. Yang K. Li 《The Journal of Horticultural Science and Biotechnology》2016,91(2):203-209
Papaya (Carica papaya L.) fruit have a short shelf-life due to rapid ripening induced by ethylene, which usually results in a high percentage of product loss. However, little is known about the genetic mechanism of ripening and the attributes of fruit quality. Ubiquitin (UBQ) proteins have received increasing attention because they play important roles in response to ripening and in regulating certain developmental processes in plants. In the present study, three genes encoding UBQ proteins, CpUBI1, CpUBI2, and CpUBI3, were isolated from papaya fruit. The lengths of the cDNAs of CpUBI1, CpUBI2, and CpUBI3 were 1,485 bp, 1,642 bp, and 529 bp, encoding 306, 308, and 156 predicted amino acids, respectively. Sequence analysis demonstrated that the CpUBI1 and CpUBI2 proteins contained four consecutive structural domains of the UBQ superfamily, while CpUBI3 contained a ribosomal domain structure of the S27a superfamily. RT-qPCR analysis demonstrated that ethephon treatment increased CpUBI gene expression significantly, compared to 1-methylcyclopropene (1-MCP) treatment and the untreated controls. Levels of CpUBI1 and CpUBI2 gene expression were significantly higher than CpUBI3. These results suggested that CpUBI1, CpUBI2, and CpUBI3 might have different roles during papaya fruit ripening and softening. 相似文献
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Quality and volatile attributes of attached and detached ‘Pluk Mai Lie’ papaya during fruit ripening
The ‘Pluk Mai Lie’ papaya (Carica papaya L.) is a promising cultivated fruit for use in fresh and processed products due to its firm flesh, but the aroma it releases is flat. Changes in quality and volatile profiles were analyzed during on- and off-tree fruit ripening. Detached fruit ripened faster than attached fruit, accumulating high internal ethylene levels. Aside from peel color, which was redder in on-tree ripened fruit, most quality attributes changed similarly in the two ripening situations. A slight increase in total soluble solids (TSS) was measured from the onset of the preclimacteric stage, whereas titratable acidity (TA) remained stable throughout the development. Whereas 2-ethyl-1-hexanol was found specifically in green fruit, ethyl octanoate emerged only in fully-ripe fruit. Furthermore, benzyl isothiocyanate was the most abundant volatile and was present in fruit at every stage except full ripening. The production of total esters, highly correlated with a loss of firmness and an increase in cavity ethylene accumulation, was about 10-fold higher in off-tree ripened fruit. The levels of methanol and ethanol sources in fruit increased steadily throughout ripening, with esters formed from ethyl alcohol predominating from the half-ripe through the senescence phases. The alcohol dehydrogenase activity in the mesocarp increased dramatically during the early ripening stages, whereas alcohol acetyltransferase was active throughout ripening. No difference in volatile profiles was found in the papaya fruit during on-tree and postharvest ripening. 相似文献
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Marcos V. Vázquez-Hernández Lourdes Arévalo-Galarza David Jaen-Contreras José L. Escamilla-García Antonio Mora-Aguilera Elías Hernández-Castro Juan Cibrián-Tovar Daniel Téliz-Ortiz 《Scientia Horticulturae》2011
The effect of inoculating ‘Maradol’ papaya plants with arbuscular mycorrhizal fungi (AMF) Glomus mosseae (GM) and Entrophospora colombiana (EC) was assessed. The results showed that both mycorrhizae species increased the number of fruits and yield in papaya plants by 41.9 and 105.2% for GM and 22.1 and 44.1% for EC, respectively, with respect to control plants. GM significantly increased plant height. Sugar content, firmness, color (°Hue), and ripening process of mycorrhized plant fruits were similar to those of the control. Weight loss of mycorrhized plant fruits was considerably less than that of the control. Inoculation of papaya with AMF is recommended, particularly with GM since it increases yield, and fruit weight (45.1%), furthermore, it reduced fruit weight loss during ripening. 相似文献
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【目的】为鉴定葡萄果实赤霉素诱导响应基因,【方法】应用cDNA-RAPD技术,以鲜食葡萄品种‘藤稔’为研究对象,对赤霉素处理后果实发育过程中基因表达进行了mRNA指纹分析。【结果】通过16条随机引物的筛选,共分离得到61个差异表达的转录衍生片段(TDF),其中增强表达或赤霉素诱导下特异性表达TDF42条,抑制表达19条。对其中18个TDF进行了克隆、测序和序列分析发现,17个TDF与NCBI已有序列同源,其中10个为已知功能基因,另有1个TDF无同源序列。对选取的6个TDF进行半定量RT-PCR验证,结果表明3个基因片段在处理条件下均特异表达或上调表达,另外3个基因片段在处理条件下均未见表达或下调表达,这与cDNA-RAPD表达谱结果相一致。【结论】利用cDNA-RAPD技术成功分离了部分受赤霉素诱导的差异表达基因。 相似文献
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W. Krongyut E. B. Esguerra J. S. Maninang S. Sugaya H. Gemma 《The Journal of Horticultural Science and Biotechnology》2013,88(6):631-639
SUMMARY‘Sunrise’ papaya fruit harvested at two stages of maturity [colour break (< 10% yellow peel colour) and 25% yellow peel colour] were treated with 100 nl l–1 1-methylcyclopropene (1-MCP) to determine its effects on ripening, on the activities and levels of gene expression of polygalacturonase (PG), pectin methyl esterase (PME), and βgalactosidase ( βGal), and on the degradation of cell wall components. 1-MCP delayed ripening and the onset of the climacteric, although the peak in the respiration rate was almost the same as that in untreated control fruit. Colour-break fruit treated with 1-MCP exhibited a continuous increase in ethylene production, but at a lower rate than in control fruit. Consequently, 1-MCP-treated fruit ripened with a concomitant reduction in firmness, which was accompanied by an increase in PG and βGal enzyme activities and gene expression. On the other hand, fruit treated with 1-MCP at the 25% yellow stage exhibited lower levels of ethylene production and developed pulp with a rubbery texture at the ripe stage which was attributed to reduced PG, βGal, and PME enzyme activities and gene expression. This was consistent with the higher level of cell wall polysaccharides measured in 1-MCP-treated fruit. The above results indicated that ‘Sunrise’ papaya fruit can be treated with 1-MCP at the colour break stage since they have a greater capacity to recover from the effects of 1-MCP than fruit treated at the 25% yellow stage. 相似文献
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Wentao Zhang Yao Tang Wei Qi Xia Liu 《The Journal of Horticultural Science and Biotechnology》2017,92(3):261-269
Apricot (Prunus armeniaca L.), a member of the Rosaceae family, with favorable nutritional and flavour quality, exhibited the characteristic climacteric changes during the fruit ripening process in ‘Shushanggan’ apricot. To further confirm the ripening-related genes in apricot, we combined quantitative proteomic tools with bioinformatics to investigate the expression profiles of ripening genes involved in significant biochemical and metabolic changes during ripening stages. The results showed that physiological traits of apricot fruits varied significantly at three different ripening stages, with remarkable correlation with fruit quality changes. Furthermore, 128 stage-specific proteins that are differently expressed in a stage-responsive manner in ripening apricot fruits were identified. Hierarchical clustering of stage-responsive proteins also revealed the metabolic changes in accordance with fruit ripening. Hence, a link has been established between protein profiles and ripening phenotypes which will help to improve our understanding of apricot fruit ripening at the proteomic level. 相似文献
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高温胁迫下番茄叶片差异表达基因的cDNA-AFLP鉴定 总被引：1，自引：1，他引：0
为鉴定番茄高温胁迫响应基因,应用cDNA-AFLP技术,以番茄耐热品系Saladette幼苗为研究对象,对高温胁迫早期叶片基因表达进行了mRNA指纹分析。通过768对引物组合的筛选,共分离得到187个差异表达的转录衍生片段(TDF),其中增强表达或高温胁迫下特异性表达TDF153条,抑制表达34条。对其中47个TDF进行了克隆、测序和序列分析。结果表明：34个TDF与NCBI或TIGR已有序列同源（E-Value<10-5）,功能涉及信号传导、转录调控以及逆境诱导蛋白等,另有13个TDF无同源序列或同源性低,预测是一些未知功能基因。 相似文献