β-半乳糖苷酶基因在猕猴桃果实成熟过程的表达

从成熟中华猕猴桃果实中克隆到了一个 β 半乳糖苷酶基因cDNA片段 ,其长度为 747bp ,有一由 2 49个氨基酸组成的开放阅读框架 ,它与苹果、芦笋、绿花椰菜、番茄中 β 半乳糖苷酶基因cDNA相应区段的核苷酸同源性为 6 7.3 %～ 70 .3 %,氨基酸同源性为6 9.1%～ 72 .7%。用该片段为探针进行Northern分析表明 ,果实采收时 ,β 半乳糖苷酶mRNA水平最高 ,随后呈下降变化 ,同时 β 半乳糖苷酶基因的表达可为外源乙烯所诱导 ,但在果实乙烯跃变期间 β 半乳糖苷酶基因的表达信号无显著变化。文中对 β 半乳糖苷酶在猕猴桃果实成熟衰老过程的作用进行了讨论     

低温对梨(Pyrus communis L.)果实后熟过程中乙烯合成和反应的影响(英文)

Passe Crassane梨果实采后需经过 6 0～ 80d的低温处理才能正常后熟。为了明确低温促进果实成熟的机理 ,对果实进行了低温和低温结合 1 MCP(1 甲基环丙烯 ,乙烯作用抑制剂 )和丙烯 (乙烯类似物 )处理。研究发现 :果实经低温处理后 ,乙烯合成前体———ACC含量大幅度升高 ,而未经低温处理的果实 ,无论贮藏在空气中或用丙烯 1 0 0 0μl/L处理 ,果实中ACC、M ACC含量均保持较低水平。但冷藏前用 1 MCP处理可抑制冷藏果实或冷藏后升温的果实ACC含量的增高。这说明果实的后熟过程与低温和依赖乙烯的ACC合成酶的活性和基因的表达密切相关。未经冷藏的果实于 2 0℃下用丙烯处理 ,果实不能自发合成乙烯 ,但当果实经过冷藏后再用丙烯处理 ,则果实对丙烯的反应能力随冷藏时间延长而增强。为了进一步了解低温诱导的乙烯反应过程。我们对乙烯受体基因进行了研究。定量PCR分析结果表明 ,与拟南芥ETR1同源的基因的表达不受低温的调节。但冷藏后升温 ,或在升温后用丙烯处理时 ,mRNA含量降低。这些结果说明 ,低温可能是通过影响乙烯信号转导途径下游的其它因子而调节依赖乙烯的ETR1基因和ACC合成酶基因的表达 ,从而影响果实的成熟过程     

甜瓜乙烯受体基因Cm-ETR1 cDNA的克隆及表达特性分析

乙烯感知和信号转导的初始成分是乙烯受体,为探明甜瓜乙烯受体基因Cm-ETR1在甜瓜果实成熟过程中的作用,以甜瓜品种河套蜜瓜为材料,根据GenBank中登录的甜瓜乙烯受体基因Cm-ETR1的cDNA序列(登录号为AF054806),设计合成特异性引物,采用RT-PCR技术克隆得到Cm-ETR1基因全长cDNA序列,提交到GenBank中(登录号为EF495185)。序列分析表明,序列长度为2 256 bp,编码区为2 223 bp,编码740个氨基酸,与已报道的cantalupenis甜瓜ETR1基因的cDNA序列完全一致。Cm-ETR1蛋白的系统进化树分析结果表明,该乙烯受体蛋白在各物种间高度保守,与黄瓜乙烯受体蛋白相似性最高,一致性为99%,与龙眼乙烯受体蛋白相似性最低,一致性为86%。定量PCR分析结果显示,随着甜瓜果实内源乙烯合成量和成熟程度的增加,Cm-ETR1基因的表达量同步增加,在果实乙烯跃变期,Cm-ETR1的表达量也达到最高值,内源乙烯合成量与Cm-ETR1基因表达量间呈显著正相关,表明Cm-ETR1基因在甜瓜果实成熟过程中可能具有重要的作用。     

乙烯受体在果实成熟及花衰老中的研究进展

乙烯受体是乙烯信号转导网络的第一个转导元件,通过调控受体基因的表达,可以调节植物对乙烯的敏感性,以调控果实的成熟及花衰老进程的响应。随着人们对乙烯受体研究的深入,乙烯受体突变体及受体抑制剂在采后果实和切花保鲜上的应用已受到广泛关注。就近年来关于乙烯受体的相关研究进展进行综述,重点介绍了乙烯受体的分子调控机制及乙烯受体在果实成熟和花衰老中的应用,并对今后乙烯受体的研究方向作了展望,以期为进一步研究提供参考。     

乙烯诱导西瓜果实的水渍化败坏

为了研究乙烯在西瓜(Citrullus lanatusThunb.Mansfeld)果实水渍化败坏过程中的作用,先将果实在5μL/L 1-甲基环丙烯(1-MCP)气体中处理18 h,然后在50 μL/L乙烯和20℃温度下贮藏.西瓜果实对乙烯处理的最初反应表现为胎座组织的电导率和游离汁液增加,同时出现组织软化和水渍化.水渍化的症状最初在靠近花萼端的内果皮中发生,在乙烯处理的第2天开始出现,ACC合成酶(ACS)和ACC氧化酶(ACO)的活性明显提高.1-MCP单独处理不产生任何明显的作用,但是会完全抑制外源乙烯诱导的水渍化败坏.没有经过乙烯处理的西瓜果实,贮藏2 d以后出现呼吸强度和乙烯释放量的高峰,10 d以后水渍化现象也零星出现.这些结果和1-MCP的预防效果说明,西瓜果实的水渍化败坏是一种由乙烯诱导的衰老现象.     

采收期对猕猴桃果实品质及其耐贮性的影响

选用海沃德猕猴桃为试验材料,对8个不同采收期(盛花后151、155、159、163、167、171、175和179 d,采收Ⅰ~Ⅷ期)猕猴桃果实在(0±1)℃冷藏过程中的生理生化变化进行比较.结果显示:(1)采收Ⅰ、Ⅱ期果实采收时具有较高的硬度、淀粉含量和可滴定酸含量,可溶性固形物含量则低于6%,贮藏期间出现的呼吸高峰和乙烯释放高峰均比其它采收期高;采收Ⅶ、Ⅷ期果实采收时硬度明显低于前面的采收期,淀粉含量和可滴定酸含量降低,可溶性固形物含量达到8%以上,贮藏期间乙烯释放速率高峰比其它采收期提前10 d,果实衰老过程明显提前;(2)入贮150 d后,采收Ⅰ、Ⅱ、Ⅶ和Ⅷ期果实的失重率差异不显著,采收Ⅰ期与采收Ⅷ期果实的腐烂率差异不显著,但均与其它采收期存在显著差异.研究发现,盛花后第159~171天采收猕猴桃,果实采收时的可溶性固形物含量在6.5%以上,贮藏120 d后与其它采收期相比仍保持相对较高的硬度、淀粉含量、Vc含量和可滴定酸含量,而且贮藏150 d后失重率和腐烂率都比较低,表明该期间为海沃德"猕猴桃的适宜采收期.     

钙对中华猕猴桃果实采后乙烯释放和呼吸的影响

不同浓度ＣａＣｌ２溶液真这渗透处理中华猕猴桃果实采后乙烯释放明显的抑制作用,其中０．５ｍｏｌ／ＬＣａ＾２＋处理尤为显著,采用５天果实人有保持极低水平的内源乙烯,乙烯峰也延迟２－４天出现。     

1-甲基环丙烯采后处理对樱桃番茄果实成熟过程的影响

研究了不同浓度(0、0.035、0.07和0.11μL/L)的乙烯受体竞争性抑制剂1-甲基环丙烯(1-MCP)采后处理对绿熟期樱桃番茄的乙烯合成、果实软化、果实色素(叶绿素、茄红素、β-胡萝卜素)含量消长的影响.0.07 μL/L及其以上浓度的1-MCP降低了前期乙烯合成,同时推迟了乙烯释放高峰,但0.035 μL/L浓度的1-MCP处理并不能抑制内源乙烯合成.1-MCP显著延迟了果实软化和叶绿素降解,但并不影响这两个过程的启动.茄红素合成的启动和积累均受到了1-MCP抑制,而1-MCP并不推迟β-胡萝卜素合成的启动,只抑制其积累.这些结果提示了乙烯调节成熟生理过程的不同机制.对于绿熟期的樱桃番茄,0.07～0.11μL/L的1-MCP是实用的有效处理浓度.1-MCP有效浓度可能用于了解果实的乙烯受体水平和乙烯敏感性.     

栀子ERF基因的克隆及其在果实成熟过程中的表达

乙烯反应元件因子(ethylene-responsive element-binding factor,ERF)是一类植物DNA结合蛋白,是乙烯信号传导过程中的一类转录因子,与植物的生长发育和生理过程有关。从栀子(Gardenia jasminoidesEllis)果实的cDNA文库中筛选获得栀子乙烯反应元件因子GjERF的cDNA。该基因全长962 bp,5’端非翻译区长82 bp,3’端非翻译区长100 bp,预测ORF为780 bp,编码259个氨基酸,分子量为28.6 kD。序列分析表明GjERF含有59个氨基酸构成的AP2/ERF结构域及N端的MC(MCGGAII)模体,它属于ERF家族的第Ⅶ亚类。RT-PCR分析表明GjERF基因在栀子成熟叶片中的表达高于在果实中的表达,并且在果实中的表达与果实的成熟过程无关。     

打孔气调贮藏与乙烯处理对猕猴桃采后品质的影响（简报）

猕猴桃(海沃德)采后果实呼吸强度和乙烯释放速率呈现明显的跃变。跃变期间果实软化,维生素C下降,总糖和还原糖上升,总酸变化不大。打孔气调贮藏有助于抑制果实软化,延缓果实衰老,延长贮藏期。采后使用100ppm乙烯处理,果实软熟加速。     

香蕉果胶裂解酶基因的克隆

根据已经报告的香蕉果胶裂解酶基因序列,设计了特异引物,通过RT-PCR获得果胶裂解酶的cDNA,并克隆测序,与已报告的序列进行了比较,二者核苷酸序列的同源性达99.24%;推测的氨基酸序列也具有很高的同源性,达97.7%.通过RT-PCR的方法对香蕉不同组织和不同成熟度果实的果胶裂解酶基因的表达进行了研究.结果表明该基因只在果实中表达,具有组织特异性,而且只在果实的特定发育阶段表达.     

乙酰水杨酸和乙烯处理对猕猴桃果实后熟软化的影响

以猕猴桃(Actinidia deliciosa(A.Chev.)C.F.Liang et A.R.Ferguson cv.Bruno)果实为试材,研究乙酰水杨酸(ASA)与乙烯处理对果实内源水杨酸(SA)含量变化以及后熟软化相关因子的影响,探讨SA在果实成熟衰老进程的作用.研究结果表明:果实后熟软化进程中,内源SA水平呈下降变化,组织中SA水平与果实硬度变化呈极显著正相关关系(r=0.969 4**),ASA处理可显著地维持组织中较高的SA水平,抑制脂氧合酶(LOX)和丙二烯氧合酶(AOS)活性增加,减低O-.2生成速率,维持细胞膜稳定性,进而抑制了乙烯生物合成或推迟乙烯跃变的到来,延缓了果实后熟软化进程,这些效应主要表现在乙烯跃变之前或乙烯跃变前期;相反,外源乙烯处理则显著降低果实组织中内源SA水平,促进LOX和AOS活性的增加,促使O-.2积累,增加了细胞膜透性,促使乙烯跃变的提前到来,加速了果实的后熟软化.推测组织中的内源SA水平与细胞膜脂过氧化作用密切相关,外源ASA可能作为一种O-.2等自由基的清除剂或是细胞膜稳定剂在组织成熟衰老过程中起作用.     

Low-temperature-modulated fruit ripening is independent of ethylene in 'Sanuki Gold' kiwifruit

Fruit ripening in response to treatments with propylene, 1-methycyclopropene (1-MCP), and low temperature was characterized in 'Sanuki Gold' kiwifruit, Actinidia chinensis Planch. Propylene treatment immediately induced rapid fruit softening, increased AC-PG (polygalacturonase) and AC-EXP (expansin) mRNA accumulation, and stimulated an increase in the soluble solid concentration (SSC) and a decrease in titratable acidity (TA). After 3?d exposure to propylene, ethylene production and AC-PL (pectate lyase) mRNA accumulation were observed. 1-MCP treatment after 24?h exposure to propylene eliminated AC-PG mRNA accumulation and suppressed continued changes in SSC and TA. Application of 1-MCP at the start of the treatment, followed by continuous propylene exposure, markedly delayed fruit softening, and the expression of the cell wall-modifying genes, and changes in the SSC and TA, indicating that kiwifruit become insensitive to ethylene at least for 3?d following 1-MCP exposure. Surprisingly, significant fruit softening, mRNA accumulation of AC-PG, AC-PL, and AC-EXP, and decreased TA were observed without ethylene production in intact fruit stored at low temperature for 1 month, but not in fruit stored at room temperature. Repeated 1-MCP treatments (twice a week) failed to inhibit the changes that occurred in low temperature storage. These observations indicate that low temperature modulates the ripening of kiwifruit in an ethylene-independent manner, suggesting that kiwifruit ripening is inducible by either ethylene or low temperature signals.     

Molecular cloning of beta-galactosidase from Japanese pear (Pyrus pyrifolia) and its gene expression with fruit ripening

We have cloned a cDNA fragment encoding a beta-galactosidase from Japanese pear (Pyrus pyrifolia) fruit (JP-GAL). It contained an untranslated sequence of 182 nucleotides at the 5' end, a presumptive coding sequence of 2,193 nucleotides and an untranslated sequence of 268 nucleotides including a polyadenylation signal and a poly (A) tail at the 3' end. It encoded a protein with a calculated molecular weight of 80.9 kDa which consists of 731 amino acids. Both the nucleotide and the deduced amino acid sequences showed a 98% sequence identity with that obtained from the apple beta-galactosidase cDNA. The peptide sequence obtained from the purified Japanese pear beta-galactosidase III matched the deduced amino acid sequence of SVSYDHKAIIINGQKRILISG (amino acid 25-45). Northern blot analysis showed that the probe derived from JP-GAL hybridized to a single 2.6 kb RNA. The mRNA was detected solely in the fruit; none was detected in the buds, leaves, roots or shoots of the Japanese pear. The steady-state level of the beta-galactosidase mRNA was measured during fruit ripening in three cultivars, Housui, Kousui (early ripening) and Niitaka (late ripening). The results showed that regardless of the cultivar, no JP-GAL mRNA was detected in the immature fruit. Increment of the mRNA level with fruit ripening coincided with the increase in the beta-galactosidase III activity. Our results showed that the expression of JP-GAL correlated with fruit softening and JP-GAL may be beta-galactosidase III.     

Characterization of ripening-regulated cDNAs and their expression in ethylene-suppressed charentais melon fruit

Charentais melons (Cucumis melo cv Reticulatus) are climacteric and undergo extremely rapid ripening. Sixteen cDNAs corresponding to mRNAs whose abundance is ripening regulated were isolated to characterize the changes in gene expression that accompany this very rapid ripening process. Sequence comparisons indicated that eight of these cDNA clones encoded proteins that have been previously characterized, with one corresponding to ACC (1-aminocyclopropane-1-carboxylic acid) oxidase, three to proteins associated with pathogen responses, two to proteins involved in sulfur amino acid biosynthesis, and two having significant homology to a seed storage protein or a yeast secretory protein. The remaining eight cDNA sequences did not reveal significant sequence similarities to previously characterized proteins. The majority of the 16 ripening-regulated cDNAs corresponded to mRNAs that were fruit specific, although three were expressed at low levels in vegetative tissues. When examined in transgenic antisense ACC oxidase melon fruit, three distinct patterns of mRNA accumulation were observed. One group of cDNAs corresponded to mRNAs whose abundance was reduced in transgenic fruit but inducible by ethylene treatment, indicating that these genes are directly regulated by ethylene. A second group of mRNAs was not significantly altered in the transgenic fruit and was unaffected by treatment with ethylene, indicating that these genes are regulated by ethylene-independent developmental cues. The third and largest group of cDNAs showed an unexpected pattern of expression, with levels of mRNA reduced in transgenic fruit and remaining low after exposure to ethylene. Regulation of this third group of genes thus appears to ethylene independent, but may be regulated by developmental cues that require ethylene at a certain stage in fruit development. The results confirm that both ethylene-dependent and ethylene-independent pathways of gene regulation coexist in climacteric fruit.     

beta-xylosidase activity and expression of a beta-xylosidase gene during strawberry fruit ripening.

Strawberry fruit shows a marked softening during ripening and the process is associated with an increment of pectin solubility and a reduction of the molecular mass of hemicelluloses. In this work, we report the activity of beta-xylosidase and the expression of a beta-xylosidase gene in strawberry fruit. We have cloned a cDNA fragment encoding a putative beta-xylosidase (FaXyl1) from a cDNA library obtained from ripe strawberry fruit. The analysis of the deduced amino acid sequence revealed that FaXyl1 is closely related to other beta-xylosidases from higher plants. The expression of FaXyl1 was strongly associated to the receptacle tissue although a low expression level was detected in achenes and ovaries. The accumulation of FaXyl1 mRNA is ripening-related, starting in white fruit, reaching the maximum at 25-50% red fruit and decreasing thereafter. The total beta-xylosidase enzyme activity was detected in all ripening stages with the maximum in 25-50% red fruit. The low activity level detected in immature stages, where no expression of FaXyl1 was found, suggests the presence of other beta-xylosidases-like genes. Both the expression of FcaXyl1 and the total beta-xylosidase activity were down regulated by auxins, as occurs for most of the ripening-related processes in strawberry fruit. A putative role of FaXyl1 and beta-xylosidase is discussed.     

Regulatory Role of Cystathionine-γ-Synthase and de novo Synthesis of Methionine in Ethylene Production during tomato Fruit Ripening

The essential amino acid methionine is a substrate for the synthesis of S-adenosyl-methionine (SAM), that donates its methyl group to numerous methylation reactions, and from which polyamines and ethylene are generated. To study the regulatory role of methionine synthesis in tomato fruit ripening, which requires a sharp increase in ethylene production, we cloned a cDNA encoding cystathionine γ-synthase (CGS) from tomato and analysed its mRNA and protein levels during tomato fruit ripening. CGS mRNA and protein levels peaked at the “turning” stage and declined as the fruit ripened. Notably, the tomato CGS mRNA level in both leaves and fruit was negatively affected by methionine feeding, a regulation that Arabidopsis, but not potato CGS mRNA is subject to. A positive correlation was found between elevated ethylene production and increased CGS mRNA levels during the ethylene burst of the climacteric ripening of tomato fruit. In addition, wounding of pericarp from tomato fruit at the mature green stage stimulated both ethylene production and CGS mRNA level. Application of exogenous methionine to pericarp of mature green fruit increased ethylene evolution, suggesting that soluble methionine may be a rate limiting metabolite for ethylene synthesis. Moreover, treatment of mature green tomato fruit with the ethylene-releasing reagent Ethephon caused an induction of CGS mRNA level, indicating that CGS gene expression is regulated by ethylene. Taken together, these results imply that in addition to recycling of the methionine moieties via the Yang pathway, operating during synthesis of ethylene, de novo synthesis of methionine may be required when high rates of ethylene production are induced.     

The Tomato E8 Gene Influences Ethylene Biosynthesis in Fruit but Not in Flowers

We investigated the function of the tomato (Lycopersicon esculentum) E8 gene. Previous experiments in which antisense suppression of E8 was used suggested that the E8 protein has a negative effect on ethylene evolution in fruit. E8 is expressed in flowers as well as in fruit, and its expression is high in anthers. We introduced a cauliflower mosaic virus 35S-E8 gene into tomato plants and obtained plants with overexpression of E8 and plants in which E8 expression was suppressed due to co-suppression. Overexpression of E8 in unripe fruit did not affect the level of ethylene evolution during fruit ripening; however, reduction of E8 protein by cosuppression did lead to elevated levels during ripening. Levels for ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC), and ACC oxidase mRNA were increased approximately 7-fold in fruit of plants with reduced E8 protein. Levels of ACC synthase 2 mRNA were increased 2.5-fold, and ACC synthase 4 mRNA was not affected. Reduction of E8 protein in anthers did not affect the accumulation of ACC or of mRNAs encoding enzymes involved in ethylene biosynthesis. Our results suggest that the product of the E8 reaction participates in feedback regulation of ethylene biosynthesis during fruit ripening.