桃果实采后软化过程中内源IAA、ABA和乙烯的变化

以玉露桃果实为试材,分析了桃果实后熟衰老进程中内源ABA、IAA和乙烯水平的变化,探讨其相互关系,以及对果实后熟软化进程的调控机制。结果表明,桃果实乙烯跃变始于采后第3天,跃变峰值出现在采后第8天;采后初期ABA含量迅速积累,于采后第3天达到高峰,随后趋下降;但IAA水平在果实后熟软化进程中呈持续下降趋势。ABA和IAA水平在后熟软化末期均有所增加。     

A label-free quantitative proteomic investigation reveals stage-responsive ripening genes in apricot fruits

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.     

ABA和IAA对猕猴桃果实成熟进程的调控

以中华猕猴桃和美味猕猴桃果实为试材,研究果实成熟过程中内源ＡＢＡ,ＩＡＡ和乙烯的变化以及外源ＡＢＡ和ＩＡＡ处理对果实后熟软化进程的调控及其在生理基础。结果表明,果实采后初期,ＡＢＡ含量迅速升高,在２－４ｄ达到最大值,之后快速下降;在ＡＢＡ下降过程中,乙烯进入跃变期,果实后熟进程加快,外源ＡＢＡ处理增加内源ＡＢＡ含量,加快内源ＩＡＡ的降解,促使脂氧合酶活性峰值提前出现,加速果实软化,果实后熟进程中,     

香蕉对甘露糖和木糖的敏感性研究

以香蕉低代试管苗茎段薄层切片和香蕉吸芽薄层切片为外植体,分别设计了13个梯度,研究了香蕉在芽的再生和愈伤组织的诱导阶段对正筛选剂D-甘露糖和D-木糖的敏感性,为建立香蕉正筛选系统的遗传转化体系奠定基础。结果表明,(1)香蕉对D-木糖敏感,在D-木糖与蔗糖比例达到10∶20时,10 d开始显示出毒性,在D-木糖与蔗糖比例达到16∶14时能显著抑制香蕉芽的再生和愈伤组织的生长,当比例达到25∶5时完全抑制香蕉组织的生长,香蕉组织彻底死亡。因此,D-木糖能作为有效筛选剂对香蕉进行目的基因遗传转化;(2)香蕉对D-甘露糖不敏感,不适用于以D-甘露糖为筛选剂的遗传转化。     

一种适合于富含多糖和酚类物质的香蕉果实RNA提取方法

酚类化合物和多糖是影响香蕉组织RNA提取的几个主要干扰因素。研究通过在裂解液中加入聚乙烯吡咯烷酮去除多酚类化合物和利用水饱和乙醚去除多糖,改进了香蕉果实RNA的提取方法。该方法具有快速、简单等特点。     

香蕉果实果胶裂解酶基因cDNA克隆及序列分析

利用已报道果胶裂解酶基因的保守序列设计简并引物,进行RT-PCR,得到1个大约1300bp的香蕉果胶裂解酶基因cDNA片段,命名为MA-pl。DNA序列分析表明:MA-pl片段全长1277bp,包含1个882bp的开放读码框(ORF),编码294个氨基酸;其具有所有果胶裂解酶共有的保守区域:钙协调部位(Asp72,Asp74,Asp96,andAsp100)、酶活性位点(Arg152,Pro154,Arg157)及3个重要的结构域(motifI:WVDH,motifII:DGLVDAVMGSTAITVSNNYF,motifIII:LYQRMPRCRHGYFHVVWNDY);MA-pl氨基酸序列与草莓-1、葡萄、草莓-2、拟南芥、苹果、香蕉(banana-1)的相似性分别为85.8%、74.2%、79.7%、78.6%、72.4%和71.4%。     

脐橙晚熟突变体及其野生型果实柠檬酸代谢基因表达分析

‘奉节晚橙’（Citrus sinensis L. Osbeck）是来源于‘奉节72-1’脐橙的晚熟芽变。采用qRT-PCR方法测定了‘奉节72-1’与其晚熟突变体果实4个发育时期柠檬酸代谢相关基因的表达。结果表明,糖酵解途径中‘奉节72-1’脐橙基因的表达高于‘奉节晚橙’;但是柠檬酸合成相关基因CsCS4、CsPEPC1、CsPEPC2、CsPEPC4、CsME1、CsME4的表达水平低于‘奉节晚橙’。果实发育后期,‘奉节72-1’脐橙中CsACO1、CsGAD4、CsGS1等柠檬酸降解相关基因表达水平逐渐升高,且表达量高于‘奉节晚橙’。两者间柠檬酸含量的差异是代谢途径中多种基因共同作用的结果。通过相关性分析,推断CsME1、CsME4可能是影响柠檬酸含量差异的关键基因。     

Acid invertase as a serious candidate to control the balance sucrose versus (glucose + fructose) of banana fruit during ripening

Six banana varieties: 3 ‘dessert’ ones: ‘IDN 110’; ‘Kirun’; and ‘Grande Naine’, and 3 ‘cooking’ ones: ‘Galéo’; ‘Sowmuk’; and ‘French’ were used to investigate the relationship between sugar profiles and activities of sucrose-phosphate synthase (SPS, EC 2.4.1.14), sucrose synthase (SuSy, EC 2.4.1.13) working in the hydrolytic way, invertases (EC 3.2.1.26) (neutral (NIV) and acid (AIV)). Expression of a Musa cell-wall invertase (MaCwINV1/pBANUU103) gene was additionally studied during fruit development and ripening ex planta after ethylene treatment of two of these varieties. Close amounts of soluble sugars (sucrose + glucose + fructose) were measured in the different varieties at ripe stage. SPS activity was either almost constant or increased continuously or transitorily during ripening of all varieties, concomitantly to total soluble sugar (sucrose + glucose + fructose) accumulation. Together with previous data obtained from ‘Cavendish’, our data lead us, (i) to strengthen the hypothesis that this enzyme is likely to have a major role in the synthesis of sucrose during ripening of banana and (ii) to underline the complexity of the mode of SPS activity regulation already pointed out. Interestingly, for the first time in banana, two diploid and cooking varieties: ‘Galéo’ and ‘Sowmuk’ were found almost sucrose-free, in good agreement with a 6.4-fold higher mostly vacuolar AIV activity than that of the two desserts and diploid ones. Otherwise, expression of a MaCwINV1/pBANUU103 (cell wall) gene was followed in the two most contrasted varieties in matter of sucrose content: ‘Sowmuk’ almost sucrose-free, and ‘IDN 110’ accumulating high level of sucrose. Compared to ‘IDN110’, the mRNA level of MaCwINV1/pBANUU103 gene was higher (up to 173-fold) in ‘Sowmuk’ concomitantly with the low level of sucrose of ‘Sowmuk’. Our data let us to conclude that the AIV is probably one of the main determinants involved in the regulation of sucrose level during banana fruit ripening, even if the form, vacuolar or cell wall-linked is not determined yet. Otherwise, the MaCwINV1/pBANUU103 gene appears as a putative candidate gene that could contribute to regulate this level.     

Responses of banana fruit to pro-long coating at different times relative to the initiation of ripening

Coating banana fruit with Pro-long 24 h after initiation of ripening decreased their skin permeability to gases and depressed their O₂ content. Coating the fruit immediately before ripening initiation delayed the onset of rapid ethylene production, which normally begins as the fruit start to ripen. Coating slightly suppressed rapid C₂H₄ production when applied immediately after ripening initiation, and exerted a strong, temporary inhibition of C₂H₄ production when applied 24 h later. A climacteric rise in respiration was absent from fruit coated with Pro-long immediately before or after ripening initiation; delaying coating by a further 24 h arrested development of the climacteric in mid-rise. In mid-climacteric fruit, inhibition of C₂H₄ production by coating occurred more rapidly than inhibition of respiration. Coating the fruit delayed the chlorophyll loss which normally accompanies ripening, but the magnitude of this effect declined as the coating treatment was delayed relative to ripening initiation. The effects of coating on skin colour change and on respiration appeared to be largely independent of its effect on the fruit's C₂H₄ content.     

主要果树病毒实时荧光定量PCR检测技术研究进展

综述了苹果、梨、柑橘、葡萄和香蕉病毒的实时荧光定量PCR检测技术研究进展,并对今后主要研究方向进行了讨论和展望。     

基于VAR模型的北京市西瓜市场的替代与整合

西瓜是我国重要的鲜活农产品之一。利用北京市批发市场中香蕉、梨子、哈密瓜、菠萝4种典型水果的价格数据,建立向量自回归模型探究香蕉、梨子、哈密瓜、菠萝4种水果在水果市场对西瓜的替代作用和动态影响,进而分析各种水果对西瓜的替代效果与水果市场的整合效应。结果表明:在不考虑季节因素的影响时,哈密瓜、香蕉、梨子与西瓜之间存在着较强的替代关系,强弱依次为哈密瓜>香蕉>梨子,而菠萝对西瓜不具有明显的替代作用。当价格受到正向的外部冲击时,西瓜自身价格会上涨,以分担这种压力,哈密瓜、香蕉和梨子这些存在替代关系的水果对西瓜价格分担上涨压力的贡献率比没有替代关系的水果高。在推动北京市西瓜市场发展的同时也要顾全整体水果市场的大局,充分考虑到各种水果之间的相互影响的替代关系。     

MaCDPK7, a calcium-dependent protein kinase gene from banana is involved in fruit ripening and temperature stress responses

Calcium-dependent protein kinase (CDPK) is an important Ca²⁺ sensor in plant development and responses to stress stimuli. Banana fruit is a typical climacteric- and chilling-sensitive fruit. The roles of CDPK genes in the ripening and chilling response of banana fruit are unclear. We isolated a cDNA fragment with full-length coding MaCDPK7 (HM061075) from fruit peel tissue. Induction of MaCDPK7 expression in fruit peel was observed 0.5 h after phytohormone ethylene treatment, earlier than the up-regulation of MaACO1 and MaACS1, coding a 1-aminocyclopropane-1-carboxylate oxidase and 1-aminocyclopropane-1-carboxylate synthase, respectively. Penetration of calcium signaling blockers, EGTA, or LaCl₃ inhibited the ripening and gene expression of MaCDPK7, MaACO1, and MaACS1 in the in vitro cultured peel pieces, but Ca²⁺ application removed the inhibitory effect of EGTA and LaCl₃. This suggested that MaCDPK7 might be a positive regulator involved in the calcium signaling in banana fruit ripening. Under temperature stresses, we found that MaCDPK7 gene expression increased 3 h after hot water dipping (HWD). The HWD-treated fruits exhibited markedly less chilling injury (CI) than control fruits in cold storage. Stored at 7°C (CI temperature) dramatically increased MaCDPK7 gene expression, while pre-treatment of HWD repressed the induction in cold storage. These results show that the MaCDPK7 gene is involved in regulating banana fruit ripening and chilling resistance induced by heat treatment.     

1-甲基环丙烯对高温下香蕉果实后熟的影响

以香蕉果实为试材,研究了1-甲基环丙烯(1-MCP)对高温逆境下香蕉果实后熟生理的影响。结果表明:35℃高温下贮藏的香蕉果实出现了明显的青皮熟现象,而0.5μL/L的1-MCP处理24h后可显著抑制高温下贮藏果实硬度的降低,延缓果皮细胞膜透性的升高,同时有效地延迟了果肉中淀粉酶活性升高、淀粉含量下降及可溶性糖含量上升。1-MCP处理果实于35℃下贮藏9d后移入20℃环境,进一步延缓了这些后熟生理变化。1-MCP和高温均抑制了果皮叶绿素含量下降,因此1-MCP处理减轻了香蕉热害的程度,延长了果实在高温下的贮藏寿命。     

香蕉泛素结合酶基因与果实成熟关系的研究

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。     

不同中间砧对苹果果实苹果酸代谢关键酶活性及其相关基因表达的影响

为深入探究中间砧影响苹果果实苹果酸代谢的机理,以SH40实生后代（代号53、111和236）为中间砧嫁接的‘天红2号’苹果树为试材,测定果实发育过程中苹果酸含量、相关代谢酶活性及基因相对表达量。结果表明：果实成熟时,以53号为中间砧嫁接的‘天红2号’果实苹果酸含量显著高于以111号为中间砧的。盛花后30、40、100 ~ 160 d,以53号为中间砧的果实中苹果酸脱氢酶（NAD-MDH）活性显著高于以111号为中间砧的;盛花后30、40和130 d,53号处理的磷酸烯醇式丙酮酸羧化酶（PEPC）活性显著高于111号处理的;盛花后100 d,以111号为中间砧的果实中苹果酸酶（NADP-ME）活性显著高于以53号为中间砧的。基因表达结果显示,盛花后100和160 d,以53号为中间砧的NAD-MDH基因相对表达量显著高于以111号为中间砧的,同时NADP-ME基因表达量也显著高于以111号为中间砧的;盛花后30 和160 d,以53号为中间砧的PEPC基因相对表达量显著高于以111号为中间砧的。     

冬枣两个ACC氧化酶基因的cDNA克隆及其表达模式

 根据植物ACC氧化酶（ACO）家族的氨基酸和核苷酸保守区序列设计简并引物,采用3′RACE技术从半红期冬枣果实中分离了两个ACO同源基因片段,随后通过PCR技术进一步获得了两个包含完整开放阅读框（ORF）及3′端非编码区（UTR）序列的ACO基因的cDNA,即ZjACO1和ZjACO2。两个基因序列在GenBank中的登录号为EU216549和EU216550,其大小分别为1 115 bp和1 105 bp,编码蛋白大小分别为319个和321个氨基酸。ZjACO1和ZjACO2在核苷酸和氨基酸水平上的同源性分别为79.4%和84.0%。Northern杂交结果表明,这两个基因在冬枣叶片中不表达,而在不同发育阶段果实中的表达具有明显差异。     

INA诱导的香蕉果实抗病性与早期活性氧积累的关系

 研究2,6–二氯异烟酸（INA）对采后香蕉果实抗病性的诱导作用和早期果皮活性氧含量、抗病相关酶活性及基因表达量的变化。结果表明：香蕉果实经0.5 g · L-1 INA处理后0、3、6、12、24 h接种炭疽病菌孢子,贮藏8 d后,INA处理果实的病斑直径比对照果实的明显减小;INA处理果实的H2O2含量和NADPH氧化酶活性分别在6 h和3 ~ 12 h明显高于对照;过氧化氢酶（CAT）活性均明显低于对照,抗坏血酸氧化酶（APX）活性稍低于对照,CAT和APX基因的表达也受到一定的抑制;内切几丁质酶（CHI）活性在0、3、6、12 h高于对照,外切CHI活性在24 h内逐步增加,而对照保持不变;CHI1和CHI2基因的表达在前6 h略高于对照。综上所述,INA诱导采后香蕉果实抗病性增加与其早期活性氧水平及抗病相关酶活性增加有密切关系,由于活性氧合成酶活性增加、清除酶活性减少而产生的高水平活性氧可能作为信号分子启动了抗病应答反应。     

Respiration and ripening patterns in the life cycle of the mango fruit

Alphonso mangoes picked at any stage of maturity (starting from fruit-set) and stored at room temperature show typical respiratory changes characteristic of a fully matured mango. Fruits picked during the earlier and later stages of development show respiration climacteric within 6–10 days. During the middle stages of development the preclimacteric trough continues to extend and the climacteric occurs after 10 days. Fully mature, tree-ripe mangoes do not show a respiratory climacteric during post-harvest storage.

Chemical constituents, such as titratable acids, apparent ascorbic acid, carbohydrates and carotenoids, estimated both at harvest and in ripe fruits from different pickings show changes similar to those characteristic of a fully matured ripening fruit.

Possibilities of reducing losses from pre-harvest fruit drop and post-harvest decay by resorting to early harvesting are discussed.     

Low temperature induce differential expression genes in banana fruits

Differential display was used to identify gene expression of banana fruit in response to low temperature stress. Banana fruits were kept at 10 °C for 8 h and 60 differential expressed fragments in pulp and peel were obtained. These fragments had an average size of 200 pb or greater and Dot blotting hybridization as well as Northern blot corroborated specific expression of these differential expressed fragments. Among the several differentially expressed genes, we found genes involved in response to pathogen attack, wounding and a ripening-associated gene. We consider these genes to belong to a general pathway which is activated upon general stress signaling.