果蝇蛋白质组学研究进展

蛋白质组学旨在阐明基因组所表达的真正执行生命活动的全部蛋白质的表达规律和生物功能。随着人类基因组学计划的逐渐成熟,分子水平的实验技术不断发展,蛋白质组学的研究被提高到了前所未有的高度。果蝇是生命科学领域最为常用的一种模式生物,长期的系统研究也使果蝇的基因组成为至今注释最好的基因组之一,为功能基因组研究奠定了基础。但由于技术的限制,迄今有关果蝇蛋白质组学研究的报道尚不多见。近年来果蝇蛋白质组学的研究主要包括表达谱、修饰谱、比较蛋白质组学和疾病模型蛋白质组等四个方向,为进一步开展人类疾病临床蛋白质组学研究奠定了基础。     

癌症差异蛋白质组学研究中样品分离和鉴定分析技术

随着人类基因组测序的完成,癌症研究的重点从基因组学转移到蛋白质组学研究中。癌症研究中的差异蛋白质组学技术也飞速发展,包括癌症样品制备、分离,蛋白质鉴定分析、蛋白质组定量研究和翻译后修饰研究等。这些技术极大地推动了与癌症相关的差异蛋白质组学研究,使蛋白质组学在癌症早期诊断、治疗,监测以及发现新药物治疗靶标方面发挥更大的作用。本文主要综述了近年来癌症差异蛋白质组学研究中样品分离和鉴定分析技术。     

生物质谱技术在蛋白质组学研究中的应用

随着技术的进步,蛋白质组学的研究重心由最初旨在鉴定细胞或组织内基因组所表达的全部蛋白质转移到从整个蛋白质组水平上阐述包括蛋白翻译后修饰、生物大分子相互作用等反映蛋白质功能的层次。多种质谱离子化技术的突破使质谱技术成为蛋白质组学研究必不可少的手段。质谱技术联合蛋白质组学多角度、深层次探索生命系统分子本质成为现阶段生命科学研究领域的主旋律之一。本文简要综述了肽和蛋白质等生物大分子质谱分析的原理、方式和应用,并对其发展前景做出展望。     

蛋白质组学的产生及其重要意义

蛋白质组学是在后基因组时代出现的一个新的研究领域,它是对机体或组织或细胞的全部蛋白质的表达和功能模式进行研究。本文简要介绍蛋白质组学产生的科学背景及其重要意义。     

植物蛋白质组学研究进展Ⅰ. 蛋白质组关键技术

随着模式植物拟南芥和水稻基因组测序相继完成, 使植物基因组学研究成功迈入到功能基因组学研究的时代。这为蛋白质组学产生及其发展奠定了坚实的基础。文章重点介绍了蛋白质组学的概念、产生背景和蛋白质组学的关键技术。蛋白质组学的关键技术包括双向电泳、高效液相色谱、蛋白芯片、质谱技术、蛋白质组学的相关数据库、定量蛋白组技术、蛋白复合体标签亲和纯化技术和酵母双杂交系统。同时对当前蛋白质组技术面临的挑战和发展前景进行了讨论。     

叶绿体的蛋白质组学研究进展

蛋白质组学是以基因组编码的所有蛋白为研究对象,高通量地从细胞及整体水平上研究蛋白质的组成及其功能的新兴学科。在后基因组时代的今天,蛋白质组学的研究正逐渐深入到生命科学的各个领域,21世纪蛋白质组学将成为生命科学中最热门的学科。蛋白质组分析已成为鉴定植物功能的有力工具之一,叶绿体作为比较重要的细胞器,在植物蛋白质组学中已有较多的研究,,随着双向电泳技术的改进和质谱法的出现,并与不断增多的拟南芥、水稻、玉米等植物的序列数据相结合,叶绿体蛋白质组可以被快速鉴定。本文主要介绍了植物蛋白质组学、叶绿体及其蛋白质组学研究技术和研究进展,并对蛋白质组学的研究趋势进行了展望。     

蛋白质组技术在小麦品质研究中的应用

人类基因组草图测定的完成,宣告了“后基因组时代”的到来。功能基因组学成为研究的重心,蛋白质组学的研究受到了空前的关注。介绍了蛋白质组研究技术的基本原理,全面综述了蛋白质组研究技术在小麦品质研究中的应用进展,包括：麦谷蛋白亚基的鉴定、面团流变学特性的遗传改良、籽粒发育过程中面筋蛋白的表达和累积、高温胁迫对面筋品质的影响、籽粒硬度蛋白分析及淀粉品质研究等,还分析了蛋白质组技术在小麦遗传育种应用研究中存在的问题与发展前景。     

昆虫差异蛋白质组: 进展和展望

黑腹果蝇Drosophila melanogaster、家蚕Bombyx mori、意大利蜜蜂Apis mellifera和冈比亚按蚊Anopheles gambiae等昆虫的基因组测序已经基本完成,蛋白质组学技术将是阐明这些基因组功能的重要工具。本文综述了应用差异蛋白质组学技术在昆虫诱导性免疫、抗性机制和分子病理研究方面取得的一些成果：（1） 细菌、真菌、脂多糖或机械损伤诱导的昆虫血淋巴或血细胞来源细胞系的蛋白质表达的改变; （2） Bt抗性昆虫中肠刷状缘膜和抗锥虫采采蝇Glossina morsitans唾液腺多种蛋白质表达的改变; （3）化学农药处理和姬蜂Chelonus inanitus携带的多分DNA病毒引起的昆虫蛋白质组的变化。最后讨论了昆虫差异蛋白质组学研究的瓶颈与对策。     

肿瘤: 一种蛋白质组病

恶性肿瘤的发生是一个涉及多因素、多基因的多阶段病理过程. 以往的研究主要集中在基因组和转录组分析. 随着人类基因组计划的完成, 肿瘤研究开始进入“后基因组时代”, 肿瘤蛋白质组学应运而生. 蛋白质作为基因功能的主要执行者, 一方面在肿瘤发生发展过程中扮演重要角色, 另一方面在很大程度上决定正常细胞和肿瘤细胞之间的差异(如异型性、恶性特征等). 本文提出, 肿瘤是一种蛋白质组病, 并根据肿瘤比较蛋白质组、表达蛋白质组、功能蛋白质组和结构蛋白质组研究进展, 从肿瘤发生发展过程中蛋白质(组)表达水平及状态、蛋白质翻译后修饰、蛋白质相互作用及网络调控等三个方面进行阐述. 该观点的提出, 将为肿瘤发病机制的研究开辟新的领域, 为肿瘤的诊断(如生物标志物筛选)和筛选(如药物新靶标)提供新的思路, 具有重要的科学意义和临床应用价值.     

蛋白质组与蛋白质组学

１９９４年澳大利有针对后基因组时代研究趋势,提出了蛋白质组和蛋白质组学的新概念。即把基因组所编码的所有蛋白为研究对象,直接探讨基因、蛋白的功能。其核心技术包括双向电泳、质谱分析、生物信息学等。该技术在探索疾病发生,寻找新药等方面取得越来越广泛的应用。     

A Deep Proteomics Perspective Into Grape Berry Quality Traits During Ripening

Discovery‐based proteomics studies have an important role in the understanding of the biochemical processes that occur during grape berry ripening. The ripening process is relevant in determining grape berry quality. For a proteome analysis of grape berry ripening, Kambiranda et al. (2018) applied a label‐free mass spectrometry–based quantitative approach. The authors reported the identification of proteins associated with the production flavor, aroma and ethylene production. Despite the valuable contribution of discovery‐based proteomics studies, the picture is still incomplete. Future efforts in gaining proteome coverage would benefit the identification of proteins associated with grape berry quality traits.     

Sequence analysis and transcriptional profiling of two vacuolar H+-pyrophosphatase isoforms in Vitis vinifera

Gene expression of grapevine vacuolar H(+)-pyrophosphatase (V-PPase EC 3.6.1.1.) during fruit ripening has previously been reported. Here we report on putative multiple V-PPase isoforms in grapevine. In this study a full-length cDNA sequence with an open reading frame of 2,295 nucleotides encoding a V-PPase gene (vpp2: acc. nr. AJ557256) was cloned. Sequence analyses of the deduced amino acid residues and RT-PCR experiments indicated that Vitis vinifera L. has at least two distinct isoforms of the V-PPase gene. Bioinformatic analyses of 13 V-PPase protein sequences revealed two highly conserved motifs associated with pyrophosphate (PPi) binding and response to stress, respectively. Both V-PPase isoforms were expressed at higher levels in the late post-véraison stage of grape berry ripening. Results also showed that the expression of grapevine V-PPase was induced by cold stress.     

Identification of grapevine aquaporins and expression analysis in developing berries

Aquaporins are membrane water channels that play critical roles in controlling the water content of cells and tissues. In this work, nine full-length cDNAs encoding putative aquaporins were isolated from grape berry cDNA libraries. A phylogenetic analysis conducted with 28 aquaporin genes identified in the grapevine genome and previously characterized aquaporins from Arabidopsis indicates that three cDNAs encode putative tonoplast aquaporins (TIPs) whereas six cDNAs belong to the plasma membrane aquaporin subfamily (PIPs). Specific probes designed on the 3' untranslated regions of each cDNA were used for the preparation of cDNA macroarray filters and in situ hybridization experiments. Macroarray data indicate that expression levels of most TIP and PIP genes depend on grape berry developmental stages and point out to a global decrease of aquaporin gene expression during berry ripening. In young berries, high expression of aquaporin genes was preferentially observed in dividing and elongating cells and in cells involved in water and solutes transport. Taken together, the data provided in this paper indicate that aquaporins are implicated in various physiological aspects of grape berry development.     

日灼对酿酒葡萄‘霞多丽’果实品质与解剖结构的影响

以酿酒葡萄品种‘霞多丽’为试验材料,采用石蜡切片与CFDA荧光染色观察日灼果皮细胞结构与细胞活性变化,同时测定日灼发生后葡萄果实品质及相关生理指标变化,以揭示日灼对果实品质与细胞结构的影响。结果表明:(1)随着葡萄日灼病加重,果实表皮颜色由浅黄色逐渐加深,后期甚至出现细胞坏死。(2)果实发生日灼后,果实硬度与含水量下降,细胞壁含量增加,果皮从外向内第1~3层细胞明显变小,细胞壁增厚。(3)随着日灼病加重,果皮细胞破裂,且破裂数量增加,细胞活性也随之下降,果皮保护功能逐渐丧失,果肉细胞逐渐失水导致了果实皱缩;重度日灼果实周缘维管束木质部导管受到果皮细胞失水断裂的影响,出现断裂变形。(4)在葡萄果实日灼发生过程中,受到高温与强光照胁迫影响,同时伴随着水分散失增加,果实可溶性固形物含量和总糖含量增加,但有机酸含量降低,糖酸比随之增加;以上各品质指标值的大小与果实水分含量密切相关。研究发现,日灼引起了葡萄果实结构变化与生理代谢的紊乱,随着日灼程度加重,果皮细胞逐渐死亡,果实内水分大量散失,果实糖含量增加,严重影响了葡萄果实的外观和内在品质。     

Proanthocyanidin synthesis and expression of genes encoding leucoanthocyanidin reductase and anthocyanidin reductase in developing grape berries and grapevine leaves

Proanthocyanidins (PAs), also called condensed tannins, can protect plants against herbivores and are important quality components of many fruits. Two enzymes, leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR), can produce the flavan-3-ol monomers required for formation of PA polymers. We isolated and functionally characterized genes encoding both enzymes from grapevine (Vitis vinifera L. cv Shiraz). ANR was encoded by a single gene, but we found two highly related genes encoding LAR. We measured PA content and expression of genes encoding ANR, LAR, and leucoanthocyanidin dioxygenase in grape berries during development and in grapevine leaves, which accumulated PA throughout leaf expansion. Grape flowers had high levels of PA, and accumulation continued in skin and seeds from fruit set until the onset of ripening. VvANR was expressed throughout early flower and berry development, with expression increasing after fertilization. It was expressed in berry skin and seeds until the onset of ripening, and in expanding leaves. The genes encoding LAR were expressed in developing fruit, particularly in seeds, but had low expression in leaves. The two LAR genes had different patterns of expression in skin and seeds. During grape ripening, PA levels decreased in both skin and seeds, and expression of genes encoding ANR and LAR were no longer detected. The results indicate that PA accumulation occurs early in grape development and is completed when ripening starts. Both ANR and LAR contribute to PA synthesis in fruit, and the tissue and temporal-specific regulation of the genes encoding ANR and LAR determines PA accumulation and composition during grape berry development.     

Xenia and metaxenia in grapes: differences in berry and seed characteristics of maternal grape cv. ‘Narince’ (Vitis vinifera L.) as influenced by different pollen sources

Literature investigations indicate that the grapes have quite complex fertilisation biology. This complexity necessitates extensive investigations to obtain reliable knowledge for both well‐organised hybridisation studies and maximising grape yield. Therefore, this study was conducted to investigate the influences of self‐, free‐ and cross‐pollination on berry and seed characteristics in grape. Five different pollination treatments were applied to ‘Narince’, the most widely known and popular white wine grape in Turkey. Pollen tests indicated that all the cultivars had satisfactory in vitro pollen viability percentages. Free‐pollination produced a significantly higher percentage berry set. Among the pollinizers, the use of pollen of ‘Thompson Seedless’ and ‘Cardinal’ varieties resulted in higher berry set percentage in ‘Narince’. The free‐pollination was also superior in giving the highest weight, length and width of the berry, as well as number of seeds per berry. These findings revealed that there were strong xenial and metaxenial effects in the studied grape cultivars. Among the pollinizer cultivars, the most effective pollinator was ‘Thompson Seedless’. Hence, for better berry set and quality, the use of ‘Thompson Seedless’ as a pollinizer may be an attractive option in both grape production and breeding studies.     

Status and future of disease protection and grape berry quality alteration by micro‐organisms in viticulture

Grapevine is one of the most widely grown fruit crops in the world. At present, however, there is much concern regarding chemical pollution in viticulture due to the application of chemical fungicides and fertilizers. One viticultural practice to resolve this issue is the application of micro‐organisms to grapevine as a substitute for chemicals. Some micro‐organisms act as an enhancer of grape berry quality as well as a suppresser of disease in grapevine through their antagonistic ability and/or systemic resistance inducing ability. Herein, we review current and prospective applications of micro‐organisms in viticulture.

Significance and Impact of the Study

In this review, we evaluate the applicability of micro‐organisms in viticulture. Micro‐organisms can improve grape berry quality through grapevine disease protection and grape berry quality alteration. Because the use of micro‐organisms to protect grapevine from plant diseases is safer than the use of chemical fungicides, the use of biofungicides in viticulture is expected to be enhanced by the increasing consumer concern towards chemical fungicides. Micro‐organisms also modify plant secondary metabolites for use as flavours, pharmaceuticals and food additives. Studies of micro‐organisms that promote polyphenol, anthocyanin and aroma compound biosynthesis are in progress with an eye to improving grape berry quality.