山葡萄糖酸积累规律及代谢调控机理研究

发布时间：2018-01-10 11:04

  本文关键词：山葡萄糖酸积累规律及代谢调控机理研究　出处：《西北农林科技大学》2016年博士论文　论文类型：学位论文

  更多相关文章： 山葡萄 光合荧光特性 生理特性 糖和有机酸代谢相关酶 代谢物运输

【摘要】：糖和有机酸含量是决定葡萄品质及葡萄酒质量的关键因素。在实际生产当中,可通过采用不同的栽培措施来调节葡萄生长的微环境、改善葡萄品质,从而获得优质的葡萄酒。本研究以集安产区的山葡萄品种北冰红和双红作为试验试材,对其光合荧光特性以及浆果内糖和有机酸含量及其相关代谢酶进行了深入的研究,并采用不同架式、不同负载和不同光呼吸抑制剂处理来研究不同栽培措施对山葡萄浆果内糖和有机酸代谢及含量的相关影响。旨在为提高当地山葡萄浆果品质和改善山葡萄生产管理模式提供科学的理论支持。主要取得以下结果:1.对山葡萄叶片光合及荧光相关指标的变化研究发现:随着山葡萄的生长发育,浆果膨大期及转色期山葡萄叶片的净光合速率(Pn)值均呈现上升态势,而始熟期山葡萄叶片的Pn呈略微下降趋势。浆果膨大期和转色期的气孔导度(Gs)日变化趋势较为接近,且三个关键时期的蒸腾速率(Tr)变化趋势与Gs几乎一致。浆果膨大期的呼吸速率(Pr)值保持最高水平,而始熟期的Pr值最低。山葡萄在成熟期时的胞间CO2浓度(Ci)值明显高于其余两个时期,而浆果膨大期和转色期的Ci值差异不大。在山葡萄浆果发育前期叶片的最大荧光(Fm)日变化较为波动,但成熟期的Fm日变化相对比较平稳。浆果膨大期山葡萄叶片在接近傍晚时仍在进行高程度的电子传递;成熟期的Fm值却略有下降,虽略有下降但未下降到最低值,表明此时光系统Ⅱ(PSⅡ)反应中心完全关闭的荧光产量仍较高,其电子传递水平也较高。PSⅡ的最大光化学效率(Fv/Fm)与PSⅡ的潜在活性(Fv/Fo)比值在不同时刻出现急剧降低表明,此刻山葡萄叶片中的PSⅡ受到了伤害,光合作用原初反应过程受抑制和影响。2.对转色期和成熟期山葡萄浆果韧皮部非离体糖卸载研究体系及韧皮部后糖运输途径的研究发现:无论是转色期还是成熟期,山葡萄非离体卸载体系浆果同化物糖分的输入在正午之前均保持较高活性,主要积累的糖分是葡萄糖和果糖;蔗糖在这两个阶段均存在,并呈缓慢下降趋势。转色期和成熟期对山葡萄浆果组织进行抑制剂处理,经“新浆果杯”法及高效液相色谱法(HPLC)法测定浆果内的糖组分及含量以及对完整葡萄浆果同化物输入动态和非离体浆果同化物卸载动态进行分析,揭示出山葡萄浆果韧皮部后糖运输的途径主要以质外体卸载途径为主,其限速关键时期主要为转色期和成熟期。3.山葡萄浆果内糖代谢相关酶活性及其糖分含量动态变化的研究发现:在整个生长季,两种架式(立架和棚架)山葡萄浆果的蔗糖磷酸合成酶(SPS)和合成方向的蔗糖磷酸(SS-s)活性均远远低于酶酸性转化酶(SAI)和SS-s的活性,且随果实发育进程而增强。从生理角度来看,较低的sps和ss-s活性和较高的sai和ss-c活性可能是成熟山葡萄浆果含糖量低的主要原因,且架式对糖代谢相关酶的影响不明显。4.对山葡萄浆果发育过程中有机酸代谢相关酶研究及有机酸含量动态变化研究发现:棚架和立架山葡萄浆果在成熟期cs活性均比浆果膨大期高。立架和棚架的pepc变化趋势近乎一致,除成熟期外,其余各个时期各个时间段内棚架的pepc酶活性均比立架模式高。两种架式的nad-idh活性变化差异十分显著,其成熟期活性均高于浆果膨大期。转色期是nad-mdh活性变化的转折点,在此之前所有棚架山葡萄浆果的nad-mdh活性均高于立架,而之后则相反。nadp-me的活性呈现先增长后下降趋势,转折点是转色中期,整个nadp-me活性在该时间点达到峰值,同时所有棚架山葡萄浆果的nadp-me活性均高于立架模式。除浆果膨大期和成熟前期,其余时间段立架mit-aco的活性均高于棚架。成熟前期是cyt-aco活性变化的转折点,在此之前,立架的cyt-aco活性均高于棚架。酒石酸和苹果酸在山葡萄整个生长季中的变化趋势较为相似。柠檬酸含量的变化趋势相对不够稳定,出现略微波动。不同架式下其柠檬酸含量的变化也存在差异。琥珀酸含量的变化无固定模式可循,两种不同架式其各自琥珀酸的含量也不尽相同。5.新型闭合y型架式对山葡萄浆果内糖分组成及糖代谢相关酶影响的研究表明:在生长发育过程尤其是转色期至成熟期内vsp和y型架式山葡萄浆果中的代谢酶sps、ss-c、ss-s及sai活性之间和浆果内葡萄糖、果糖、蔗糖和乳糖含量之间存在一定差异。架式对葡萄糖和果糖的影响主要集中在转色期之后直到成熟期,但对乳糖含量并无太大影响。vsp架式对蔗糖含量的影响主要集中在成熟期而y型架式对蔗糖含量的影响主要集中在转色中期至成熟期。转色前期,vsp和y型架式能够通过调节sps和ss-c的活性进而调节蔗糖的浓度;转色后期,ss-c活性的轻微增加而sai的活性未受影响。转色后期,两种架式中的ss-s和sps都能够调节其他糖分,如葡萄糖、半乳糖、麦芽糖、阿拉伯糖及果糖。6.不同光呼吸抑制剂nahso3和inh对山葡萄光合特性及单糖(ms)和二糖(ds)含量影响的研究表明:三种浓度(150、250、350mg/l)的nahso3处理均对山葡萄光呼吸有强烈抑制作用并能显著提高光合产物(可溶性固形物ss,ms+ds)的产量及品质,且效果优于inh的三个处理,并以250mg/l浓度表现最优。inh能够提高山葡萄叶片的光合荧光特性,但对光合产量的提高效果不明显。7.不同负载对山葡萄浆果糖和有机酸代谢影响的研究发现:山葡萄品种北冰红和双红均属低蔗糖类型。二者的蔗糖含量在转色前期最高,随后在浆果发育过程中逐渐被消耗直到浆果完全成熟。除成熟期时双红高负载处理外,两种品种各处理间浆果内果糖的含量变化与葡萄糖含量的变化几乎一致。中负载处理的两种山葡萄品种均含有各自最高的葡萄糖及果糖含量。不同负载对山葡萄浆果内有机酸有较大影响:酒石酸和苹果酸含量随着浆果的成熟而逐渐降低,两种品种中负载处理在成熟期时均含有各自最低含量的柠檬酸、苹果酸以及酒石酸;北冰红和双红在各时期,其浆果内的苹果酸含量均高于酒石酸含量,且中负载的苹果酸含量低于低负载和高负载处理。不同负载对柠檬酸和琥珀酸含量并无明显影响。综合分析可知,无论是北冰红还是双红山葡萄品种,中负载处理是得到浆果内最优糖酸含量的最佳选择。
[Abstract]:Sugar and organic acid content is a key factor in determining the quality of grape and Wine quality. In actual production, micro environment can be adjusted by using different grape cultivation measures for growth, improve grape quality, so as to obtain high quality Wine. In this study, in Anchan mountain grape varieties Beibinghong and double red as test material, the photosynthetic characteristics and the berries in sugar and organic acid content and related metabolic enzymes were studied, and the different type, different loads and different respiratory inhibitors treatment on different cultivation measures on correlation of grape berry sugar and organic acid metabolism and to provide support for the content. The scientific theory to improve the local grape berry quality and improve the grape production management model. The main results are as follows: 1. of grape leaf photosynthesis and fluorescence correlation index change The study found that: with the grape growth, net photosynthesis during berry expansion period and mountain grape leaf color of photosynthetic rate (Pn) value showed a rising trend, while the beginning maturity grape leaf Pn showed a slightly decreasing trend. Berry expansion period and stomatal conductance of veraising (Gs day) the trend is more close, and the transpiration rate of three key periods (Tr) change trend with Gs is almost the same. Berry expansion period respiration rate (Pr) value to maintain the highest level, and initial maturity stage of the lowest Pr. Grape maturity in intercellular CO2 concentration (Ci) value was significantly higher than the other the two period, and berry enlargement stage and color changing of Ci value had little difference. In the early development of grape berry fluorescence maximum leaf (Fm) diurnal variation more volatile, but Fm diurnal variation of maturity is relatively smooth. Berry expansion period mountain grape leaves near dusk is still in the high degree of electronic Transfer; mature Fm value declined slightly, although a slight decline but did not decrease to the minimum value, show that this time system II (PS II) fluorescence yield completely closed reaction center is still high, the electron transfer will have a higher level of maximum photochemical efficiency of.PS (Fv/Fm) and potential activity of PS II (Fv/Fo) the ratio in different time decreases sharply at the moment that grape leaves in PS II was hurt by the inhibition of photosynthetic primary reaction process and the effect of.2. on the color turning stage and mature stage of grape berry phloem in vitro non sugar unloading research system and Postphloem sugar transport pathways were found that the color change stage or maturity, non isolated mountain grape berries assimilation sugar unloading system input and maintain a high activity before noon, the main accumulation of sugar is glucose and fructose; sucrose in these two stages are kept in, and slowly down Drop trend. Color turning stage and mature stage of inhibitor treatment on grape berries, the "new berry Cup" method and high performance liquid chromatography (HPLC) method for the determination of berry sugar composition and content of grape berry complete assimilation input dynamic and non isolated berry assimilate unloading dynamics analysis revealed from grape berry Postphloem sugar transport pathway mainly in the apoplastic unloading pathway, the rate limiting critical period is mainly veraison and maturity of.3. glucose metabolism related enzyme activity in grape berries and dynamic changes of sugar content found in the whole growth season, the two type (Li frame and scaffolding) sucrose phosphate synthase grape berries (SPS) and synthesis direction of sucrose phosphate (SS-s) enzyme activity were far lower than the acid invertase (SAI) and the activity of SS-s, and increased with fruit development process from the physical point of view, Sai and SS-C activity and lower SPS and higher ss-s activity may be the main reason of mature grape berries with low sugar content, and the influence of posture on enzymes related to sugar metabolism was.4. on organic acid metabolism in grape berry development of enzyme and organic acid content dynamic change research found: scaffolding and stand mountain grape berries in CS activity than mature berry expansion period high. The change trend of PEPC stand and the scaffolding was almost the same, in addition to mature stage, PEPC activity in the rest of each period of each period of time are high than scaffolding frame model. The difference of NAD-IDH activity changes of two kinds of posture is very significant, its maturity the activity was higher than that of veraison berry expansion period. The change of nad-mdh activity is a turning point, before all the scaffolding mountain grape berry nad-mdh activity was higher than the vertical frame, and then the opposite.Nadp-me activity showed first increase After the decline, the turning point is the mid turn color, the activity of nadp-me at the time point and reached the peak at the same time, all scaffolding Vitis amurensis berry nadp-me activity were higher than that of vertical frame mode. In addition to Berry expansion period and mature period, the remaining time of the activity of mit-aco was higher than that of vertical frame scaffolding. Early maturing period is the turning point, the activity of cyt-aco change prior to this, the activity of cyt-aco was higher than that of vertical frame scaffolding. Tartaric acid and malic acid are similar in the whole growth trend of grape Ji Zhong. The change trend of citric acid content is not stable, slightly fluctuations. Different changes in the type of citric acid content is also different. The changes of the content of succinic acid fixed pattern, the research content of two kinds of different type of their respective succinic acid are not the same type.5. type Y type closure related enzyme effect on grape berry sugar composition and glucose metabolism Show: SS-C in the process of growth and development especially metabolizing enzymes SPS, colour changed stage to mature stage of VSP and Y type mountain in grape berries, glucose, ss-s and Sai activity and berries in fructose, there are some differences between the sucrose and lactose content. Effects of type of glucose and fructose are mainly concentrated in the turn after color until maturity, but the content of lactose have little effect on the effect of.Vsp type sucrose content mainly concentrated in the mature period and the effect of Y type on sucrose content mainly concentrated in the middle turn color to maturity. Color stage, the concentration of VSP and Y type frame type can through regulating SPS and SS-C activity and then adjust the sucrose; color changing period, slightly increased SS-C activity and Sai activity were not affected. Color change and SPS two ss-s later, the rack can be adjusted to other sugars, such as glucose, galactose and maltose, glucose and fructose in Arabia .6. and INH on photorespiration inhibitor NaHSO3 grape photosynthetic characteristics and monosaccharides (MS) and two (DS) of sugar content effects showed that three concentration (150250350mg/l) of the NaHSO3 treatments on grape photorespiration have an inhibitory effect and can significantly improve the photosynthetic products (soluble solids SS, ms+ds) the yield and quality, and the three treatment effect is better than that of INH, and the concentration of 250mg/l.Inh can be best to improve grape leaf fluorescence light, but photosynthetic yield increasing effect is not obvious.7. different load on mountain grape pulp fructose and affect the metabolism of organic acid. Mountain grape varieties Beibinghong and double red are low sucrose type. The sucrose content of the two highest early in the color turning, then during berry development is gradually consumed until berries fully mature. In addition to the mature red double high processing load, two varieties of each treatment The content change of change and the content of glucose in fructose between berries are nearly same. Two kinds of grape varieties of processing load contain their highest glucose and fructose content. Different load has a great influence on the grape berries in organic acid, tartaric acid and malic acid content decreased with the fruit mature, two kinds of load the treatment of citric acid contained the lowest content in their maturity varieties, malic acid and tartaric acid; Beibinghong and double red in each period, the content of malic acid in the berries were higher than tartaric acid content, low load and high load is lower than the content of malic acid load. And different load effect on citric acid and succinic acid. There is no comprehensive analysis shows that, whether Beibinghong or double Hongshan grape varieties, in the processing load is the best choice to get the optimal berry sugar and acid content.

【学位授予单位】：西北农林科技大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S663.1
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本文编号：1405004