早红考密斯及其芽变果实中花青素含量与相关酶活性变化的研究

以西洋梨早红考密斯及其绿色芽变果实为材料,研究了果实发育期间果皮色泽、花青苷含量及其相关酶活性变化.结果显示:(1)早红考密斯果皮色泽从成熟前的暗红色逐渐变为成熟时的浅红色,并在色泽分布不均匀的地方显出黄色底色,色泽指数(a*)值从花后45 d的16.4降低到成熟时的7.4,降低54.9%;花青苷含量从成熟前的258.4μg?g-1降到成熟时的118.3μg?g-1;早红考密斯果皮色泽和果皮花青苷含量具有密切的相关性.(2)早红考密斯的绿色芽变在果实发育的前期检测不到花青苷,发育后期果实向阳部出现浅红晕,但花青苷含量极低,与亲本差异极显著.(3)果实发育期间,两品种间苯丙氨酸解氨酶(PAL)变化趋势相似,总体呈下降趋势,且早红考密斯的活性总体低于其绿色芽变;两品种查耳酮异构酶(CHI)活性总体变化趋势基本一致,均呈现缓慢上升的趋势,在前期绿色芽变的CHI活性高于其亲本,后期低于亲本;类黄酮3-O-葡(萄)糖基转移酶(UFGT)活性在两品种间的差异较大,在整个果实发育期间早红考密斯的UFGT活性远高于其绿色芽变.研究表明,早红考密斯果皮色泽变化主要由花青苷的含量不同引起;PAL和CHI不是绿色芽变的直接原因;UFGT与花青苷合成密切相关,绿色芽变果皮中UFGT活性显著降低.     

‘赛蜜酥1号’枣及其芽变品系果实的性状差异北大核心CSCD

该试验以‘赛蜜酥1号’枣及其芽变品系‘赛蜜酥2号’果实为试验材料,运用流式细胞术对它们的倍性进行鉴定,采用石蜡切片法进行果实细胞组学的观察,并对两者果实生长发育过程中的内外观品质及决定果实口感的蔗糖代谢相关酶活性进行比较分析,为深入研究芽变对枣果实口感、品质造成的影响提供理论支持。结果表明:(1)‘赛蜜酥2号’芽变品系的细胞倍性未发生改变,仍为二倍体。(2)两个枣品种(系)果实在外部形态上有明显区别,‘赛蜜酥1号’为卵圆型,‘赛蜜酥2号’为扁圆型,且后者的果形指数成熟时大于1,单果质量高于前者,核小、可食率更高,皮薄果实更加酥脆;‘赛蜜酥1号’果皮的蜡质层厚度一直大于其芽变枣,两者角质层厚度和表皮层厚度的变化趋势基本一致,但厚度之间具有显著差异。(3)影响果实细胞生长、分裂,果实脱落的各类激素水平在品种间均存在显著差异,‘赛蜜酥2号’IAA和GA_(3)含量显著高于‘赛蜜酥1号’,使之果形更大。(4)两个枣品种(系)果实可溶性糖、可滴定酸、维生素C、植物淀粉、可溶性蛋白含量等随发育时期的变化趋势一致,但含量水平之间存在差异,‘赛蜜酥2号’果实的糖含量、维生素C含量更高,可滴定酸含量更低,口感更甘甜。(5)两品种枣果实的蔗糖代谢相关酶活性在果实迅速生长的膨大期都存在着显著差异,也导致‘赛蜜酥2号’果实甜度明显高于‘赛蜜酥1号’。研究发现,两个枣品种均为二倍体,它们在果实外部形态上存在明显区别,易于分辨;芽变品系‘赛蜜酥2号’果实更大,核小、可食率更高,皮薄更加酥脆,口感更甘甜。     

橄榄果实发育成熟过程中糖积累与相关酶活性的关系

以可溶性总糖含量差异明显的2个橄榄品种为试验材料,测定果实发育成熟过程中蔗糖、葡萄糖、果糖、可溶性总糖含量及蔗糖代谢相关酶活性的动态变化,并对果实糖积累与酶活性进行相关性分析,以明确不同橄榄品种果实糖积累差异的生理基础,为进一步在代谢与分子水平探讨橄榄果实糖积累机制提供依据。结果表明:(1)蔗糖快速积累期是橄榄品种间果实蔗糖积累差异的关键时期,并影响成熟时果实可溶性总糖含量的高低,其中‘马坑22’蔗糖快速积累期较长,增长幅度较大,成熟时可溶性糖含量高;成熟时‘马坑22’、‘檀头23’果实内己糖与蔗糖比分别为0.668、0.904。(2)在蔗糖快速积累期内,‘马坑22’酸性转化酶(AI)活性低于‘檀头23’,为其蔗糖积累创造条件,而中性转化酶活性高于后者则有利于其增加果实库强;两品种蔗糖磷酸合成酶(SPS)活性变化差异不大,说明SPS不是蔗糖积累的关键酶;‘马坑22’蔗糖合成酶(SuSy)合成方向活性在花后144~186d增幅显著高于‘檀头23’,说明SuSy为果实蔗糖积累的关键酶。(3)‘马坑22’蔗糖快速积累主要依靠SuSy合成方向活性变化促进蔗糖合成,‘檀头23’蔗糖快速积累主要依靠SuSy分解方向活性变化促进蔗糖直接进入果实。     

杨梅果实发育进程中的碳水化合物代谢

以‘乌紫’和‘荸荠’两个杨梅品种为试材,测定了干鲜重、糖含量、可滴定酸含量、蔗糖和己糖代谢相关酶活性的动态变化。结果表明,杨梅果实的干鲜重、含糖量的快速增长和可滴定酸含量的快速下降均发生在果实发育后期。成熟‘乌紫’杨梅果实的蔗糖含量约占总糖的2／3以上,而‘荸荠’杨梅仅为总糖的49％。‘荸荠’杨梅的转化酶和蔗糖合酶分解活性随着果实发育呈上升趋势,‘乌紫’杨梅的则变化不大。两个品种的蔗糖磷酸合酶活性随着果实发育呈上升趋势,但蔗糖合酶合成活性到果实发育中期后下降。两个品种的己糖激酶活性变化相似,但果糖激酶活性的变化趋势不同。     

不同梨品种果实有机酸含量变化与相关酶活性的研究

以梨品种‘鸭梨’、‘茌梨’和‘八里香’为试材,对不同品种果实有机酸积累及相关酶活性随发育的动态变化特征进行分析比较,探讨果实有机酸含量与相关酶活性的关系。结果表明:(1)梨果实发育过程中有机酸含量呈逐渐下降趋势,果实成熟时的总有机酸含量以‘茌梨’最高,‘鸭梨’次之,‘八里香’最低,且3品种间总酸含量差异显著。(2)果实发育后期,‘鸭梨’、‘茌梨’和‘八里香’的柠檬酸含量几乎相同,而苹果酸的含量差异显著,说明苹果酸是引起品种间总酸含量差异的主要因素。(3)在苹果酸代谢中,不同品种间NADP-ME酶活性在果实发育后期差异显著,而NAD-MDH和PEPC酶活性无显著差异,表明NADP-ME酶是引起‘鸭梨’、‘茌梨’和‘八里香’成熟果实苹果酸含量差异的主要原因。(4)不同品种的柠檬酸代谢酶CS、Cyt-ACO、Mit-ACO和NAD-IDH的活性变化趋势一致,且成熟时无显著差异,表明它们不是引起梨果实酸含量差异的主要原因。(5)梨果实发育过程中有机酸含量与相关代谢酶活性的相关性分析表明,苹果酸和柠檬酸代谢酶共同影响梨果实的酸含量水平。     

桃果实成熟前后细胞壁成分和降解酶活性的变化及其与果实硬度的关系

比较桃品种‘双久红’和‘川中岛白桃’果实成熟前后20 d内果肉硬度、细胞壁成分和细胞壁降解酶活性变化的结果表明,桃果实成熟5 d后,‘双久红’桃果实的硬度、纤维素含量和原果胶含量均极显著高于‘川中岛白桃’:从成熟前15 d开始,‘双久红’的水溶性果胶含量、多聚半乳糖醛酸酶活性和纤维素酶活性均极显著低于‘川中岛白桃’;整个成熟期间,‘双久红’的果胶甲酯酶活性明显低于‘川中岛白桃’。相关分析表明,果实硬度与原果胶、纤维素含量呈极显著正相关,而与可溶性果胶含量、多聚半乳糖醛酸酶活性和纤维素酶活性呈极显著负相关。     

‘京红’桃芽变缝合线局部早熟分子机制的探究北大核心CSCD

为了探究桃缝合线局部早熟的分子机制,该研究以‘京红’桃芽变(JHM)及其野生型(JHW)的果实为试材,测定分析了缝合线和果面部位的硬度、花青素含量以及差异基因的表达特征。结果显示:(1)‘京红’桃芽变比其野生型果实晚成熟约2周,芽变果实缝合线部位比果面部位局部早成熟,且提前2周时间转为红色。(2)随着果实的成熟,‘京红’桃野生型及其芽变的果实硬度逐渐降低,花青素含量逐渐升高,并均在花后66 d发生明显变化,芽变缝合线部位硬度比果面部位更低,花青素含量比果面更高。(3)在花后66 d,芽变果实的缝合线与果面部位差异表达基因数高达1889个,显著富集在代谢途径、次生代谢产物的生物合成、植物激素信号转导、苯丙素的生物合成等代谢途径;从中筛选到24个缝合线早熟相关基因,包含5个细胞壁降解相关基因,9个色素合成、调控相关基因,5个乙烯合成与转导相关基因,3个生长素应答基因和2个NAC转录因子。(4)对24个早熟相关基因中的12个差异表达基因进行荧光定量验证结果表明,基因表达趋势与转录组测序结果相一致。研究发现,桃芽变果实种仁产生的乙烯通过缝合线向周围扩散,促进缝合线部位ACS1和ACO1等基因的转录,并合成了较多乙烯,乙烯又进一步调控该部位PG、XTH33、CHS、DFR等细胞壁降解与色素合成相关基因的表达,导致该部位的果肉提前成熟。     

硬肉桃果实成熟前后钙含量和钙ATP酶活性的变化及其与果实硬度的关系

以硬肉桃新品种‘双久红’果实为试材,以常规优良品种‘川中岛白桃’为对照,分别研究了成熟前20d和成熟后20d内两品种果实中钙含量和Ca^2＋-ATP酶活性变化以及它们与果肉硬度关系的结果表明：‘双久红’果实的总钙和Ca^2＋含量从成熟前15d开始极显著高于同期‘川中岛白桃’的,两者与果实硬度变化呈极显著相关（P〈0．01）,随着果实的成熟两者均呈下降趋势,Ca^2＋-ATP酶的调控能力也逐渐减弱,但‘双久红’果实中的Ca^2＋-ATP酶活性比‘川中岛白桃’高一些。     

‘南红梨’采后生理特性及其后熟过程中的品质变化分析

以‘南果梨’及其红色芽变品种‘南红梨’为试材,对两种果实的采后生理特性及后熟期间品质变化进行比较分析。结果表明:(1)与‘南果梨’相比,‘南红梨’果实外观红色鲜艳,可溶性固形物含量高、可滴定酸含量低、固酸比较高,口感更甜。(2)采后‘南红梨’呼吸高峰值及后熟前期乙烯生成量显著高于‘南果梨’,果实硬度下降更快,并在贮藏第6天至第18天均小于同期的‘南果梨’果实,使口感更绵软。(3)最佳食用期时,两种果实均检测到43种香气成分,其中40种相同,并各有3种香气成分对方为检出;‘南红梨’香气成分总生成量显著大于‘南果梨’,并且酯类香气成分相对含量(80.335%)显著高于‘南果梨’(75.713%),因此,‘南红梨’的香气更加浓郁。     

中华猕猴桃和毛花猕猴桃果实碳水化合物及维生素Ｃ的动态变化研究

分别对中华猕猴桃（Actinidia chinensis）黄肉品种‘金桃’和毛花猕猴桃（Actinidia eriantha）品系‘6113’果实生长发育过程中碳水化合物及维生素C的动态变化进行了系统研究。结果表明,中华猕猴桃‘金桃’和毛花猕猴桃‘6113’果实的可溶性固形物（SSC）含量均于谢花后146d内保持相对平稳,而后开始上升;此时,两物种果实的淀粉含量均上升到最大值,之后两者均开始下降。两者糖含量的变化与SSC相似,且中华猕猴桃‘金桃’果实糖含量进入快速增长期的时间比毛花猕猴桃‘6113’早1个月。两者果实Vc含量的变化趋势相似,均于7月上中旬达到一个高峰,以后随着果实的生长发育,含量下降,‘金桃’于8月14日降至最低值,‘6113’于9月13日降至最低值;两者的Vc含量降到最低值后均缓慢上升,到果实完全成熟期（树上自然软熟期）回升到第二个峰值。‘6113’果实的Vc含量在完全成熟期的峰值远远高于7月上旬的高峰值。对‘金桃’和‘6113’果实碳水化合物及Vc含量方差分析表明,两者的可溶性固形物、淀粉和总糖没有明显差异,而毛花猕猴桃‘6113’的Vc含量显著高于中华猕猴桃‘金桃’。     

根瘤蚜对不同抗性葡萄的选择性及葡萄根系挥发性物质的鉴定

为了探讨不同抗性水平砧木及品种对根瘤蚜吸引及驱避效应以及鉴定挥发性物质组分,采用离体根段培养法测定葡萄根瘤蚜Phylloxera viticola Fitch对砧木5BB,140Ru及栽培品种巨峰的选择性,发现根瘤蚜对栽培品种巨峰的根段具有很强的选择性,至接种后3 d,67.13％的根瘤蚜优先选择巨峰;而对5BB和140Ru的选择性却很差,分别有15.29％和17.58％的根瘤蚜选择5BB及140Ru。根瘤蚜在不同品种根段上的生长发育和繁殖也存在明显差异。对砧木5BB和巨峰根系的挥发性组分进行测定,发现2个品种挥发物独有组分数量均较高,巨峰独有组分为23个,占总成分数的48.94％,5BB的独有组分为32个,占总成分数的57.14％。5BB和巨峰的共有组分为24个,主要成分均为亚油酸甲酯和反油酸甲酯,但含量有所不同,其中,5BB以倍半萜类的丁香醇及脂肪酸甲酯类的亚油酸甲酯,反油酸甲酯,棕榈油酸甲酯含量较高,高于巨峰中相应物质含量的4.23%~6.46％,而巨峰以邻苯二甲酸二丁酯及胆甾烷含量较高,分别高于5BB相应组分含量的6.81％和1.07％。结果说明葡萄根瘤蚜对不同抗性水平砧木及品种确实存在不同的选择性,并且不同抗性水平的砧木及品种的挥发性物质存在显著差异。     

Developmental changes in the diurnal water budget of the grape berry exposed to water deficits

The diurnal water budget of developing grape (Vitis vinifera L.) berries was evaluated before and after the onset of fruit ripening (veraison). The diameter of individual berries of potted ‘Zinfandel’ and ‘Cabernet Sauvignon’ grapevines was measured continuously with electronic displacement transducers over 24 h periods under controlled environmental conditions, and leaf water status was determined by the pressure chamber technique. For well-watered vines, daytime contraction was much less during ripening (after veraison) than before ripening. Daytime contraction was reduced by restricting berry or shoot transpiration, with the larger effect being shoot transpiration pre-veraison and berry transpiration post-veraison. The contributions of the pedicel xylem and phloem as well as berry transpiration to the net diurnal water budget of the fruit were estimated by eliminating phloem or phloem and xylem pathways. Berry transpiration was significant and comprised the bulk of water outflow for the berry both before and after veraison. A nearly exclusive role for the xylem in water transport into the berry was evident during pre-veraison development, but the phloem was clearly dominant in the post-veraison water budget. Daytime contraction was very sensitive to plant water status before veraison but was remarkably insensitive to changes in plant water status after veraison. This transition is attributed to an increased phloem inflow and a partial discontinuity in berry xylem during ripening.     

Polyuronide solubilization during ripening of normal and mutant tomato fruit

The amount and molecular size of soluble polyuronide extractable from ripening tomatoes is markedly affected by residual enzyme activity. The efficacy of phenol-acetic acid-water treatment to remove this residual activity is demonstrated. Data obtained using treated wall preparations confirms that there is an increase in soluble polyuronide during normal ripening and that this also occurs in the ‘Never-ripe’ mutant, and to a lesser degree in the ‘ripening-inhibitor’ mutant. However, changes in the molecular size of this polyuronide during normal ripening were not as extensive as previously reported and few changes were apparent in either of the mutants.Measurements were also made of polygalacturonase (EC 3.2.1.15) and pectinesterase (EC 3.1.1.11) activity during ripening. The level of polygalacturonase activity does not appear to correlate with the amount of soluble polyuronide released, but may be related to the extent of depolymerisation. No relationship was apparent between the level of pectinesterase and either soluble polyuronide released or depolymerization.     

Changes in cell-wall polysaccharides from the mesocarp of grape berries during veraison

Softening of grape berries ( Vitis vinifera L. × V. labruscana cv. Kyoho) was evaluated by studying changes in composition and degradation of cell-wall polysaccharides. The grape berry softens at the beginning of the second growth cycle many weeks before harvest. The softening stage is called 'veraison' by viticulturists. On day 50 after full bloom, green hard berries (before veraison [BV]), softening berries (veraison [V]) and partly peel colored berries (C) were selected from the same clusters. In addition, mature berries (M) were collected on day 78 after full bloom. Mesocarp tissues at each stage were fractionated into hot water-soluble (WS), hot EDTA-soluble (pectin), alkali-soluble (hemicellulose) and residual (cellulose) fractions. Neutral and acidic sugar contents of WS and pectin fractions decreased only after the V stage, while the neutral sugar content of the hemicellulose fraction decreased from the BV to V stages. Cellulose content constantly decreased as the berry ripened, but the large decrease was found from the BV to V stages. Molecular masses of pectic and hemicellulosic polysaccharides decreased from the BV to V stages. Hemicellulosic xyloglucan was markedly depolymerized from the BV to V stages. The neutral and acidic sugar composition of each fraction changed little during the berry ripening. These data indicated that softening of berry during veraison involved the depolymerization of pectin and xyloglucan molecules and decrease in the amounts of hemicellulose and cellulose.     

Sugar accumulation in grape berries. Cloning of two putative vacuolar invertase cDNAs and their expression in grapevine tissues.

During grape berry (Vitis vinifera L.) ripening, sucrose transported from the leaves is accumulated in the berry vacuoles as glucose and fructose. To study the involvement of invertase in grape berry ripening, we have cloned two cDNAs (GIN1 and GIN2) from berries. The cDNAs encode translation products that are 62% identical to each other and both appear to be vacuolar forms of invertase. Both genes are expressed in a variety of tissues, including berries, leaves, roots, seeds, and flowers, but the two genes have distinct patterns of expression. In grape berries, hexose accumulation began 8 weeks postflowering and continued until the fruit was ripe at 16 weeks. Invertase activity increased from flowering, was maximal 8 weeks postflowering, and remained constant on a per berry basis throughout ripening. Expression of GIN1 and GIN2 in berries, which was high early in berry development, declined greatly at the commencement of hexose accumulation. The results suggest that although vacuolar invertases are involved in hexose accumulation in grape berries, the expression of the genes and the synthesis of the enzymes precedes the onset of hexose accumulation by some weeks, so other mechanisms must be involved in regulating this process.     

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.     

Physiology of the tomato mutant alcobaca

Alcobaca is commonly regarded as an abnormally ripening mutant of the tomato (Lycopersicon esculentum Mill.). Alcobaca fruits were found to be similar to cv. Rutgers fruits in the following characteristics: time between full anthesis and the onset of ripening, response to ethephon, flavor, pH and concentrations of titratable acids, total soluble solids and reducing sugars. The pattern of CO₂ and ethylene climacteric are similar in the two plant types, but the peak levels were lower and occurred later in alcobaca than in ‘Rutgers’. The mutant fruits differed from fruits of normal varieties in their greatly prolonged shelf life, their relatively low activity of polygalacturonase (PG) and polymethylgalacturonase (PMG), and their low level of endogenous ethylene. Fruits of the mutant harvested before the onset of ripening failed to reach normal pigmentation and remained yellow. Fruits harvested at the onset of ripening reached an orange color, while fruits ripened while attached to the plant reached almost normal pigmentation. These results suggest that alcobaca is a slow ripening mutant and does not belong to the category of non-ripening mutants.