葡萄叶片中营养物质和叶绿素含量与其对绿盲蝽抗性的关系

【目的】研究葡萄品种的抗性对于绿盲蝽Apolygus lucorum综合防控策略的制定具有重要意义。本研究检测了不同葡萄品种对绿盲蝽的抗性水平,分析了叶片中可溶性蛋白、可溶性糖和叶绿素含量与其对绿盲蝽抗性的关系,以明确不同葡萄品种对绿盲蝽的抗性机理。【方法】以9个葡萄品种(巨峰、玫瑰香、红地球、红宝石、藤稔、赤霞珠、蛇龙珠、霞多丽和品丽珠)为材料,通过连续3年的田间抗性鉴定确定这些葡萄品种对绿盲蝽的抗性水平;分别采用考马斯亮蓝法、蒽酮比色法和乙醇提取法测定不同葡萄品种叶片中可溶性蛋白、可溶性糖和叶绿素的含量,并通过线性回归法分析这些物质含量与葡萄对绿盲蝽抗性的关系。【结果】不同葡萄品种对绿盲蝽的抗性水平不同,按抗性指数从小到大依次为红地球、玫瑰香、巨峰、赤霞珠、品丽珠、红宝石、蛇龙珠、藤稔和霞多丽;不同葡萄品种叶片中可溶性蛋白、可溶性糖和叶绿素含量存在差异;可溶性蛋白和叶绿素含量与抗性指数之间没有显著相关性,而可溶性糖含量与抗性指数之间存在显著负相关(P0.05)。【结论】不同葡萄品种对绿盲蝽的抗性存在显著差异,藤稔和霞多丽对绿盲蝽具有较高抗性,而红地球、玫瑰香和巨峰则较为敏感,该差异可能与葡萄叶片中可溶性糖的含量有关。     

单氰胺对葡萄休眠过程中冬芽水分和碳水化合物的影响

以酿酒葡萄品种'赤霞珠'和'霞多丽'为材料,对其冬芽休眠进程和休眠过程中冬芽水分和碳水化合物含量的变化进行分析.结果表明,(1)在陕西杨陵地区气候条件下,'霞多丽'、'赤霞珠'分别在12月20日和1月9日达到深度休眠,随后开始休眠解除阶段.(2)在芽休眠过程中,两个品种冬芽的总水分含量、自由水含量和淀粉含量均随休眠的加深而降低,随休眠的解除而增加,而冬芽的束缚水含量、束缚水/自由水比值、可溶性糖含量均在休眠加深阶段持续上升,在休眠解除阶段逐渐降低.(3)单氰胺处理后,两品种冬芽中自由水含量显著增加,束缚水的比例同时显著降低,且'赤霞珠'的变化幅度大于'霞多丽',但其总水分、可溶性糖和淀粉等含量在休眠过程中无显著变化.(4)葡萄冬芽中的水分含量及存在状态、可溶性糖含量和淀粉含量的变化与其休眠进程密切相关;单氰胺处理能够增加冬芽自由水含量,降低其束缚水含量,从而有效打破葡萄休眠.     

两个葡萄杂交后代根系抗葡萄根瘤蚜及抗寒性鉴定

【目的】为了筛选抗葡萄根瘤蚜Daktulosphaira vitifoliae Fitch且抗寒的葡萄砧木以适应我国葡萄生产需求。【方法】以山葡萄Vitis amurensis Rupr.左山1号×SO4杂种F1代的45个株系(A系列)和左山1号×101-1杂种F1代27个株系(B系列)为试材,采用离体根接种鉴定法进行抗葡萄根瘤蚜鉴定及抗性分级;采用差热分析系统(differential thermal analysis,DTA)进行各株系根系的低温放热(low temperature exotherms,LTE)分析,建立各株系根系韧皮部及木质部的温度-伤害度(LT-I)回归方程,评估各株系根系的抗寒性。【结果】葡萄根瘤蚜在杂交株系根系上的产卵量均显著低于敏感品种巨峰,筛选出被葡萄根瘤蚜侵染后不能形成根瘤,抗葡萄根瘤蚜级别为0级的A系列杂交株系18个和B系列株系11个。被葡萄根瘤蚜侵染后形成根瘤比例低于10%的抗葡萄根瘤蚜级别为1级的A系列杂交株系9个和B系列株系4个;筛选出A系列综合低温放热温度隶属度函数、韧皮部和木质部低温放热温度隶属度函数3个指标均低于贝达的株系27个,B系列各指标均低于贝达的株系3个。【结论】本研究筛选出抗寒性强且对葡萄根瘤蚜抗性强的A系列株系15个和B系列株系2个。其中,A14,A16,A18,A22,A23,A28,A34,A35,A38,A44,A50,B24和B26对葡萄根瘤蚜抗性级别为0;A11,A15,A17和A27对葡萄根瘤蚜抗性级别为1。     

贺兰山东麓酿酒葡萄的生态适应性

从抗寒性、光合特性、生长及其结果性状等方面研究了宁夏贺兰山东麓酿酒葡萄的生态适应性。室内抗寒性鉴定和田间抗寒评价结果表明：‘歌海娜’最不耐寒,抗寒性强的品种有‘长相思’、‘雷司令’、‘黑比诺’、‘佳美,;抗寒力较差的品种有‘赛美容’、‘白诗南’、‘白玉霓’、‘神索’;抗寒力较强的品种是：‘霞多丽’、‘赤霞珠,、‘梅鹿辄’、‘白比诺’、‘西拉’。对酿酒葡萄的光合特性的测定发现：同一立地不同品种的净光合速率（Pn）差异明显,‘赤霞珠’的光合速率远高于‘霞多丽’、‘梅鹿辄’的,各品种光合速率的日变化呈双“S”曲线;全天光合速率的最高峰出现在上午10：00左右,下午14：00出现光合低谷,次高峰出现在下午17：00左右。同一品种在不同立地上的叶面积（LA）、叶鲜重（LFW）、比叶重（SLW）、叶绿素（Ch1）含量均表现差异;不同品种在同一立地条件下以上几个指标也表现不同。酿酒葡萄在贺兰山东麓不同区域的生长结果表现也不同。研究表明：大多数酿酒葡萄品种在贺兰山东麓具有良好的生态适应性,土壤和品种是造成贺兰山东麓酿酒葡萄生态适应性差异的主要因素,该区酿酒葡萄栽培的首要任务是解决抗寒栽培技术和土壤培肥。     

TTC染色鉴定葡萄抗寒性方法的优化与应用北大核心CSCD

以‘左山1号’、101-14、3309C、SO4、140R和‘黑比诺’等葡萄砧木和品种直径为0.5~0.7 cm的一年生枝条为材料,经低温梯度(0℃、-14℃、-19℃、-24℃、-29℃和-34℃)处理后,对TTC染色的温度和时间进行优化并观察统计葡萄枝条不同组织的存活情况,测定枝条的相对电导率,以及韧皮部和木质部的可溶性蛋白、游离脯氨酸、可溶性糖、淀粉含量和束缚水自由水比例(束自比)5个生理指标,分别用枝条纵切面染色面积、相对电导率拟合Logistic方程计算枝条的半致死温度(LT_(50))来评价枝条的抗寒性,同时通过隶属函数法综合评价枝条韧皮部和木质部抗寒性,并将3种方法的评价结果进行比较,以建立一种直观高效鉴定葡萄品种抗寒性的方法。结果表明:(1)依据低温胁迫下各品种葡萄砧木扦插枝条的萌芽率和生根率表现,其抗寒性由强到弱依次为‘左山1号’、101-14、3309C、SO4、140R和‘黑比诺’。(2)葡萄枝条TTC活力染色的最佳条件为pH=7.0,0.5%TTC-0.1 mol/L磷酸缓冲液在35℃下避光染色36 h。随着胁迫温度的降低,各品种枝条纵切面染色面积均逐渐降低,根据优化TTC法获得‘左山1号’、101-14、3309C、SO4、140R和‘黑比诺’枝条的LT_(50)分别为-31.38℃、-26.51℃、-26.10℃、-23.60℃、-23.33℃和-19.26℃。(3)随着胁迫温度的降低,各品种枝条的相对电导率逐渐增加,且‘黑比诺’的相对电导率基本保持最高、增幅最大(51.93%),而‘左山1号’的相对电导率始终最低、增幅最小(44.07%);根据相对电导率获得‘左山1号’、101-14、3309C、SO4、140R和‘黑比诺’枝条的LT_(50)分别为-30.02℃、-26.40℃、-25.75℃、-23.16℃、-21.13℃和-17.72℃。(4)通过5种生理指标进行枝条抗寒性隶属函数综合性评价结果显示,同一品种中韧皮部抗寒性强于木质部,同一部位不同品种的抗寒性表现为:‘左山1号’>101-14>3309C>SO4>140R>‘黑比诺’。研究发现,TTC染色法与电导法和综合隶属函数法获得的葡萄枝条抗寒力评价结果一致,也与枝条生长恢复鉴定结果一致;但与其他两种方法相比,TTC染色法能更加直观和有效地预测评估葡萄枝条的抗寒性。     

青杨脊虎天牛的危害与杨树氨基酸组成和含量的相关性

为了研究不同品种杨树上青杨脊虎天牛Xylotrechus rusticus L.的危害与树木氨基酸组成和含量之间的相关性,利用HPLC柱前AccQ·Tag衍生法,分析35 a小青杨、24 a 小青×黑杨、24 a 小黑杨、16 a 香杨、27 a和40 a小叶杨的韧皮部和木质部中氨基酸的组成及相对含量。在各杨树品系主干2.5 m高处,韧皮部共检测出15种氨基酸：丙氨酸、精氨酸、天冬氨酸、半胱氨酸、胱氨酸、谷氨酸、甘氨酸、异亮氨酸、亮氨酸、赖氨酸、苯丙氨酸、脯氨酸、丝氨酸、苏氨酸和组氨酸;木质部氨基酸种类与韧皮部的相近,区别在于木质部缺少组氨酸,多了缬氨酸。各杨树品系韧皮部同种氨基酸的相对含量差异显著,但与青杨脊虎天牛危害株率无紧密相关性;木质部中同种氨基酸相对含量差异显著,且与青杨脊虎天牛危害株率呈显著正相关。结果提示杨树木质部氨基酸相对含量越高,越利于天牛幼虫的生长发育。小青杨中氨基酸种类齐全、含量均衡,其株被害率高达84%;小黑杨木质部氨基酸种类和相对含量都较贫乏,香杨的氨基酸种类不全,且相对含量不均衡,二者均不受青杨脊虎天牛危害。不同部位株被害率的差异与氨基酸的相对含量无关,可能与青杨脊虎天牛只在老树皮裂缝处产卵的习性有关。     

短期环割对骆驼刺气孔导度及叶绿素荧光的影响

为研究骆驼刺对不同环割的生理响应,以塔克拉玛干沙漠南缘策勒绿洲外围骆驼刺为试验材料,设置了对照(CK),韧皮部半割(PS),韧皮部全割(PF),木质部半割(XS)4种处理,研究了骆驼刺的气孔导度、光合色素含量、Chl a/b、叶绿素荧光参数和曲线在不同环割处理下的变化特征。结果表明:短期韧皮部半割对骆驼刺各项生理指标影响不明显;韧皮部全割下,骆驼刺气孔导度、光合色素含量、叶绿素荧光参数有了不同程度的下降;木质部半割显著降低了骆驼刺气孔导度但光合色素含量和大部分荧光参数没有发生变化。处理后第5天和第10天各项参数相差不大,短期内环割对骆驼刺的伤害程度随时间推移变化较小。总体来看,短期环割处理对骆驼刺伤害程度是PFXSPSCK。在荒漠化防治过程中,应尽量避免骆驼刺的韧皮部被全部剥蚀。     

葡萄休眠解除过程中冬芽组织活性氧与抗氧化系统的变化特征

以酿酒葡萄‘赤霞珠’(红)和‘霞多丽’(白)为试材,通过考察其自然休眠过程和打破休眠(单氰胺处理)过程中冬芽组织活性氧及抗氧化系统的变化,探讨葡萄休眠机制。结果表明:(1)在陕西杨陵地区,早熟品种‘霞多丽’在12月20日左右进入深度休眠,第2年1月29日进入休眠解除阶段;晚熟品种‘赤霞珠’在1月9日左右达到深度休眠,2月18日左右进入休眠解除阶段;单氰胺处理可促进葡萄休眠的解除。(2)葡萄休眠过程中,活性氧O2.-和H2O2含量在休眠初期逐渐增加,深度休眠时含量最高,休眠解除过程中含量降低,从而促进休眠的解除;超氧化物歧化酶(SOD)活性在休眠中一直降低,休眠解除后活性上升;过氧化物酶(POD)活性在休眠前期较低,随后活性上升,休眠解除过程中活性又降低;过氧化氢酶(CAT)活性在休眠过程中持续上升,随后维持在较高水平;抗坏血酸-还原态的谷胱甘肽(ASA-GSH)循环在休眠前期较弱,休眠解除过程中逐渐增强。研究发现,葡萄进入休眠后,其冬芽抗氧化能力逐渐降低,至深度休眠时抗氧化能力降至最低,随着低温积累需冷量得到满足,活化了抗氧化系统进而抗氧化能力迅速提高,休眠被解除。     

镉胁迫下杞柳对金属元素的吸收及其根系形态构型特征

采用水培方法,研究了杞柳(Salix integra)2个品种在0-90 μmol/L Cd处理下不同组织对Cd的吸收和积累规律,探讨了镉胁迫下杞柳根系形态学功能响应特征以及对矿质离子吸收的影响。结果发现,镉在杞柳2个品种不同组织的含量均表现为根>韧皮部>木质部>叶。2个品种地上部组织对镉的吸收和积累规律相似:在0-70 μmol/L Cd处理浓度范围内,随着溶液Cd浓度的增加,叶、木质部、韧皮部中镉的含量逐渐增加,到50 μmol/L时,镉在地上部组织的含量达到最高,当Cd处理浓度达90 μmol/L时,地上部各组织中Cd含量出现下降趋势。而2个品种根系对Cd的吸收则不同,"微山湖"品种在溶液Cd达70 μmol/L时,根系镉的积累量最大,在90 μmol/L时,根系Cd的积累量明显下降;"一枝笔"品种根系在0-90 μmol/L范围内,根系Cd的积累量均为增加趋势。通过分析2个品种根系形态学参数变化发现,Cd胁迫抑制了杞柳2个品种根的伸长,促进了"微山湖"品种根的径向生长,导致根系平均直径增加;进一步对叶片矿质营养状态分析发现, Ca、Mn的吸收受镉胁迫的影响较大,在高浓度镉胁迫下(50-70 μmol/L)"微山湖"和"一枝笔"对Ca和Mn的吸收明显下降;Cd胁迫对Fe在叶片的含量影响不明显;同时发现,在50 μmol/L Cd处理下,Cu²⁺在叶片的积累明显增加。由此可见,杞柳2个品种间对镉的吸收和积累差异主要体现在根系,地上部对镉毒害的响应差异不大。     

四个酿酒葡萄品种组培快繁体系的初建

为了提高酿酒葡（Vitis vinifera）苗木繁殖速度及苗木品质,以‘赤霞珠’、‘西拉’、‘霞多丽’和‘美乐’4／p品种为试材,研究无菌外植体建立、启动培养、增殖培养和驯化移栽环节的关键技术,初步建立酿酒葡萄组培快繁体系。结果表明,以半木质化茎段为外植体接种成活率高,在培养基MS＋IBA0．2mg·L-1＋6．BA1．0mg·L-1＋KT0．5mg·L-1上启动培养外植体单芽萌发率最高,以培养基1／2MS＋IBA0．2mg·L-1＋KT1．0mg·L-1增殖培养兼生根诱导,组培苗生长健壮,繁殖率高。增殖培养6代后,‘赤霞珠’、‘西拉’、‘霞多丽’和‘美乐’分别由12株葡萄苗扩繁为l383株、1095株、744株和100株。组培苗驯化培养3周后移栽至营养钵,4个品种成活率均在72％以上。此组培快繁体系基本适用于4个酿酒葡萄品种,可应用于科学研究及工业化大规模生产。     

Dynamic thermal time model of cold hardiness for dormant grapevine buds

Background and Aims

Grapevine (Vitis spp.) cold hardiness varies dynamically throughout the dormant season, primarily in response to changes in temperature. The development and possible uses of a discrete-dynamic model of bud cold hardiness for three Vitis genotypes are described.

Methods

Iterative methods were used to optimize and evaluate model parameters by minimizing the root mean square error between observed and predicted bud hardiness, using up to 22 years of low-temperature exotherm data. Three grape cultivars were studied: Cabernet Sauvignon, Chardonnay (both V. vinifera) and Concord (V. labruscana). The model uses time steps of 1 d along with the measured daily mean air temperature to calculate the change in bud hardiness, which is then added to the hardiness from the previous day. Cultivar-dependent thermal time thresholds determine whether buds acclimate (gain hardiness) or deacclimate (lose hardiness).

Key Results

The parameterized model predicted bud hardiness for Cabernet Sauvignon and Chardonnay with an r² = 0·89 and for Concord with an r² = 0·82. Thermal time thresholds and (de-)acclimation rates changed between the early and late dormant season and were cultivar dependent but independent of each other. The timing of these changes was also unique for each cultivar. Concord achieved the greatest mid-winter hardiness but had the highest deacclimation rate, which resulted in rapid loss of hardiness in spring. Cabernet Sauvignon was least hardy, yet maintained its hardiness latest as a result of late transition to eco-dormancy, a high threshold temperature required to induce deacclimation and a low deacclimation rate.

Conclusions

A robust model of grapevine bud cold hardiness was developed that will aid in the anticipation of and response to potential injury from fluctuations in winter temperature and from extreme cold events. The model parameters that produce the best fit also permit insight into dynamic differences in hardiness among genotypes.     

Identification of indigenous yeast flora isolated from the five winegrape varieties harvested in Xiangning,China

Inoculated fermentation by selected indigenous yeast strains from a specific location could provide the wine with unique regional sensory characteristics. The identification and differentiation of local yeasts are the first step to understand the function of yeasts and develop a better strain-selection program for winemaking. The indigenous yeasts in five grape varieties, Chardonnay, Cabernet Franc, Cabernet Sauvignon, Marselan, and Merlot cultivated in Xiangning, Shanxi, China were investigated. Eight species of seven genera including Aureobasidium pullulans, Candida zemplinina, Hanseniaspora uvarum, Hanseniaspora occidentalis, Issatchenkia terricola, Metschnikowia pulcherrima, Pichia kluyveri, and Saccharomyces cerevisiae were identified using Wallerstein Laboratory Nutrient medium with sequencing of the 26S rDNA D1/D2 domain. H. uvarum and S. cerevisiae were the predominant species, while most non-Saccharomyces species were present in the whole fermentation process at different levels among the grape varieties. The genotypes of S. cerevisiae from each microvinification were determined by using interdelta sequence analysis. The 102 isolates showed eight different genotypes, and genotype III was the predominant genotype found. The distribution of S. cerevisiae strains during the fermentation of Marselan was also studied. Six genotypes were observed among the 92 strains with different genotypes of competitiveness at different sampling stages. Genotype V demonstrated the potential for organizing starter strains and avoiding inefficient fermentation. In general, this study explored the yeast species in the grapes grown in Xiangning County and provided important information of relationship of local yeast diversity and its regional wine sensory characteristics.     

Genetic Transformation of Major Wine Grape Cultivars of Vitis Vinifera L.

We have developed an Agrobacterium-mediated transformation system for a number of important grapevine cultivars used in wine production. Transgenic plants were obtained for the seven cultivars: Cabernet Sauvignon, Shiraz, Chardonnay, Riesling, Sauvignon Blanc, Chenin Blanc and Muscat Gordo Blanco. Embryogenic callus was initiated from anther filaments and genotypic differences were observed for initiation and subsequent proliferation with Chardonnay responding most favourably to culture conditions. The transformation system allowed the recovery of germinating transgenic embryos 10–12 weeks after Agrobacterium inoculation and plants within 18 weeks. Examination of the expression patterns of the green fluorescent protein gene under the control of the CAMV35S promoter in leaf tissue of transgenic plants showed that for up to 35% of plants the pattern was not uniform. The successful transformation of a genetically diverse group of wine grape cultivars indicates that the transformation system may have general application to an even wider range of Vitis vinifera cultivars.     

葡萄枝条水分含量变化与抗寒性关系

以12个葡萄品种1年生枝条为试材,通过0、-15、-18、-21、-24、-27、-30、-133℃8个低温处理24h后,测定不同低温胁迫下葡萄枝条的,6-含水量、束缚水含量、自由水含量和束缚水与自由水的比值。结果表明：随着处理温度的降低,同一品种枝条的总含水量基本恒定,自由水含量呈先下降再升高的趋势,束缚水含量和束缚水与自由水的比值均呈现为先升高再下降的趋势;束缚水与自由水的比值随低温胁迫的变化呈Cubic方程,不同品种束缚水与自由水的比值达到最高点的温度不同,‘双红’、‘北冰红’、‘左优红’的拐点温度在-25℃以下,‘775’、‘巨峰’的拐点温度在-20℃左右,‘雷司令’、‘黑比诺’、‘霞多丽’的拐点温度在-18℃左右,‘赤霞珠’、‘梅鹿特’、‘白比诺’、‘红地球’拐点温度在-16～17℃。对葡萄休眠枝条进行低温处理后,通过测定束缚水与自由水含水量变化鉴定葡萄品种的抗寒性是可行的。     

Performance and diacetyl production of commercial strains of malolactic bacteria in wine

This study compares 11 commercial cultures of Leuconostoc oenos and Lactobacillus plantarum in Cabernet Sauvignon, Pinot Noir and Chardonnay wines. Performance of the cultures was found to be greatly influenced by wine type. Better survival of the bacteria was observed in Cabernet Sauvignon and Pinot Noir wines. The time necessary to complete malolactic fermentation (MLF) was 65 ± 14 d for Chardonnay, 71 ± 3 d for Cabernet Sauvignon, and 25 ± 8 d for Pinot Noir. The maximal rate of malate utilization was 0·4 g d^-1 for Pinot Noir, and 0·2 g d^-1 for the two other wine types. Final diacetyl concentration was lower in Chardonnay wines (highest 0·58 mg l^-1) compared to the other wines (highest 5·8 mg l^-1). Malic and citric acid were co-metabolized by all strains. None of the strains metabolized glycerol. Significant differences in final diacetyl concentration of wine vinified with the different strains were found. Panelists could reliably differentiate MLF wines from non-MLF wines, irrespective of their diacetyl content, indicating that diacetyl is not the only important MLF flavour.     

Performance of several models for predicting budburst date of grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.)

The budburst stage is a key phenological stage for grapevine (Vitis vinifera L.), with large site and cultivar variability. The objective of the present work was to provide a reliable agro-meteorological model for simulating grapevine budburst occurrence all over France. The study was conducted using data from ten cultivars of grapevine (Cabernet Sauvignon, Chasselas, Chardonnay, Grenache, Merlot, Pinot Noir, Riesling, Sauvignon, Syrah, Ugni Blanc) and five locations (Bordeaux, Colmar, Angers, Montpellier, Epernay). First, we tested two commonly used models that do not take into account dormancy: growing degree days with a base temperature of 10°C (GDD₁₀), and Riou’s model (RIOU). The errors of predictions of these models ranged between 9 and 21 days. Second, a new model (BRIN) was studied relying on well-known formalisms for orchard trees and taking into account the dormancy period. The BRIN model showed better performance in predicting budburst date than previous grapevine models. Analysis of the components of BRIN formalisms (calculation of dormancy, use of hourly temperatures, base temperature) explained the better performances obtained with the BRIN model. Base temperature was the main driver, while dormancy period was not significant in simulating budburst date. For each cultivar, we provide the parameter estimates that showed the best performance for both the BRIN model and the GDD model with a base temperature of 5°C.     

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.     

Mining and validating grape (<Emphasis Type="Italic">Vitis</Emphasis> L.) ESTs to develop EST-SSR markers for genotyping and mapping

Grape expressed sequence tags (ESTs) are a new resource for developing simple sequence repeat (SSR) functional markers for genotyping and genetic mapping. An integrated pipeline including several computational tools for SSR identification and functional annotation was developed to identify 6,447 EST-SSR sequences from a total collection of 215,609 grape ESTs retrieved from NCBI. The 6,447 EST-SSRs were further reduced to 1,701 non-redundant sequences via clustering analysis, and 1,037 of them were successfully designed with primer pairs flanking the SSR motifs. From them, 150 pairs of primers were randomly selected for PCR amplification, polymorphism and heterozygosity analysis in V. vinifera cvs. Riesling and Cabernet Sauvignon, and V. rotundifolia (muscadine grape) cvs. Summit and Noble, and 145 pairs of these primers yielded PCR products. Pairwise comparisons of loci between the parents Riesling and Cabernet Sauvignon showed that 72 were homozygous in both cultivars, while 70 loci were heterozygous in at least one cultivar of the two. Muscadine parents Noble and Summit had 90 homozygous SSR loci in both parents and contained 50 heterozygous loci in at least one of the two. These EST-SSR functional markers are a useful addition for grape genotyping and genome mapping.