Effect of crop load on hardiness,protein and amino acids content of apple flower buds at the wintering stage and the begining of the growth∗

The cold tolerance of flower buds of apple Malus domestica Borkh. ('Mclntosh'/M.7) declined during spring growth. The absence of fruit in the previous year increased the ability of the buds to withstand lower temperatures. All flower buds retained the capacity to tolerate freezing injury until first pink stage. The median lethal temperature (MLT) of the flower buds increased relatively throughout the sampling period. The flower buds of cropped trees were more susceptible to low temperature injury during spring at all stages of development. The flower buds of non‐cropped trees contained higher levels of hydrophilic and acidic amino acids from October until April, which may be related to their greater cold tolerance. A new nitrogen:protein conversion factor (5.51) was calculated based on total amino acid analysis which could be substituted for 6.25 as a means of estimating total protein content of apple flower bud tissue using the Kjeldahl method.     

桃MPK3基因的克隆、表达及生物信息学分析

为探究MPK3响应GA信号调控休眠解除的分子机理,以中油四号油桃为试验材料,通过实时荧光定量PCR发现GA诱导下MAPKs家族基因在桃芽中有不同程度的表达,其中PpMPK3响应GA正调控表现最强烈。利用MEGA6.0、MEME、GSDS、DNAMAN 6.0等软件对桃PpMPK3基因进行生物信息学分析,结果表明,PpMPK3的cDNA全长1 113 bp,编码370个氨基酸,蛋白分子量约为42.6 kD,理论等电点为5.62,属于亲水性不稳定蛋白。预测PpMPK3蛋白发生磷酸化修饰的位点有45个,不存在糖基化位点。PpMPK3蛋白的二级结构中α-螺旋占41.35%,延伸链占10.54%,无规则卷曲为48.11%。进化树分析表明,PpMPK3氨基酸序列与甜樱桃亲缘关系最近,同源性为99.73%。通过亚细胞定位发现,PpMPK3定位于细胞核。RT-PCR结果表明,PpMPK3的表达模式与休眠解除过程相一致,推测PpMPK3能够响应GA信号,促进休眠解除。本研究结果为桃设施栽培的休眠调控提供了一定的理论依据。     

Tastes and structures of bitter peptide, asparagine-alanine-leucine-proline-glutamate, and its synthetic analogues

Asn-Ala-Leu-Pro-Glu (NALPE) is a strong bitter peptide with a minimum response threshold (MRT) of 0.074 mM. To elucidate the relationship of spatial structure and bitterness on peptides, NALPE and its analogues, NALPW, NALPS, NALPL, NALPP, NALPD, and NALPR, were synthesized and sensorially evaluated. Structural analysis using computer simulation for each peptide revealed that the presence of a polar group and hydrophobic bitter amino acids, the composition of hydrophobic regions, the spatial orientation of the polar group and hydrophobic regions, and the proximity between polar groups and hydrophobic regions faced within the same plane space may be the major determinants for the taste type and intensity of peptide bitterness.     

遮荫对休眠期大樱桃芽呼吸代谢的影响研究

试验研究遮荫处理对休眠期大樱桃花芽、叶芽呼吸代谢的影响结果表明 ,休眠前期大樱桃花芽和叶芽呼吸强度均较弱 ,先期缓慢下降且叶芽降幅低于花芽 ,低温来临后其呼吸强度有较大幅度下降 ,之后变化较小 ;休眠末期其呼吸强度缓慢上升。自然条件下 3条主要呼吸途径变化趋势不同 ,糖酵解途径呈下降趋势 ,三羧酸循环途径始终徘徊在 4 8%左右 ,磷酸戊糖途径则呈明显上升趋势。磷酸戊糖途径不断得到活化 ,推测其增强可能是解除大樱桃休眠的主要原因 ,这与打破种子休眠的研究结果一致。遮荫处理对大樱桃花芽和叶芽呼吸强度的影响趋势类似 ,即 10月 2 4日前其呼吸强度迅速下降 ,之后变化较小。遮荫处理使花芽呼吸强度降幅略大于叶芽 ,且遮荫程度对其影响也大于叶芽。遮荫处理对花芽和叶芽呼吸途径的影响一致 ,与对照相比遮荫处理使糖酵解途径所占比率降低 ,三羧酸循环途径比率略有增加而磷酸戊糖途径比率显著上升 ;2层遮荫处理糖酵解途径降幅和磷酸戊糖途径增幅均大于 1层遮荫处理。遮荫处理使磷酸戊糖途径不断增强 ,表明遮荫处理可能是通过呼吸代谢而促进大樱桃芽休眠的解除。     

Heavy metal binding by hydrophobic and hydrophilic dissolved organic carbon fractions in a Spodosol A and B horizon

For a one year period intact Spodosol soil columns were percolated weekly with H₂O_deion, 1.58 mmol H₂SO₄ L^?1, and 0.79 mmol H₂SO₄ L^?1+0.64 mmol HNO₃ L^?1, respectively. Decomposition rates, soil organic carbon (OC) solubilization, dissolved organic carbon (DOC) fractions, and Cr-, Cu-, and Cd-binding by dissolved hydrophobic and hydrophilic acids were studied. Acid treatment reduced significantly OC respiration as well as OC solubilization in the humic layers. The reduced OC solubility at acid addition was more pronounced for the less polar hydrophobic compounds, resulting in a decrease of the hydrophobic acids (from ca. 65 to 40–45% of DOC), and in an increase of the hydrophilic acids (from ca. 25 to 40–45% of DOC). For B horizon leachates, DOC increased at acid treatment. Generally, hydrophobic acids were retained preferentially in the B horizon. Also in the B horizon output there was an increase of the hydrophilic acids as acidity increased (from ca. 40 to 50% of DOC). Differences between the two acid treatments were negligible. The degree of metal-organic complexes decreased in the order Cr >Cu >Cd, from A to B horizon leachates, and with increasing acidity. Hydrophilic acids were found to be the dominating ligands in complexing Cr and Cu. Actual Cr- and Cu-binding by hydrophilic acids exceeded that by hydrophobic acids 2–8 times. As the hydrophilic acids represented the most mobile DOC components in the soil columns, in particular with increasing acidity, significant amounts of Cr and Cu in the B horizon leachates were organically complexed, although a great proportion of the hydrophobic acids was retained in the B horizon.     

海马水解蛋白的氨基酸组成与抗氧化能力的关系

为了探讨大海马蛋白质的高效利用方法,本试验将脱脂大海马粉经碱性蛋白酶和胰蛋白酶联合消解后,酶解液(PH)经超滤膜分离为5组分(PH-Ⅰ、PH-Ⅱ、PH-Ⅲ、PH-Ⅳ、PH-Ⅴ),结合各组分的氨基酸组成,回归分析了各组分的自由基清除能力与氨基酸组成间的相关关系。结果表明,酶解液经超滤膜分离的5组分中PH-Ⅴ(<2 500 Da)抗氧化能力较强,清除DPPH自由基能力为34.2%±0.1%,对超氧阴离子的清除能力为29.2%±0.1%,Fe³⁺还原力为0.28%±0.1%。海马酶解多肽各组成中疏水性氨基酸含量低于亲水性氨基酸,但疏水性氨基酸对自由基的清除能力起着决定性作用,极性氨基酸和非极性氨基酸可通过协同作用来增强海马多肽清除自由基的能力,且每个单位极性氨基酸的抗氧能力是非极性氨基酸的1.2倍,每个单位芳香族氨基酸的抗氧化能力是脂肪族氨基酸的2倍。本研究不仅制备了一类具有开发价值的抗氧化多肽,并且分析了多肽抗氧化能力与氨基酸组成的相关关系,为大海马的高值化利用提供了理论依据。     

茶树生长素抑制蛋白基因CsARP1的克隆与表达分析

从茶树休眠芽抑制消减杂交文库中分离得到生长素抑制蛋白基因的3＇-片段,以休眠芽为材料,利用RACE技术克隆了其cDNA全长,并利用荧光定量PCR研究了该基因在不同休眠阶段芽的相对表达量。结果从茶树休眠芽中获得一个全长为711bp的生长素抑制蛋白基因CsARP1（GenBank登录号为HQ225758）。该基因开放阅读框...     

间伐改形对陇东密闭苹果园树体枝条和花芽及果实品质的影响

为研究间伐改形对陇东成龄乔化密闭富士苹果园树体枝类组成、花芽形成及果实品质的影响,以16年生密闭红富士苹果园为研究对象,对隔株间伐改形和不间伐改形树体枝类组成、花芽形成、质量和果实品质等指标进行比较分析。结果表明,间伐改形后长枝和中枝占比率比不间伐改形分别减少5.97、5.88百分点;短枝占比率和优质短枝占比率比不间伐改形分别增加11.85、24.97百分点;顶花芽量比不间伐改形提高35.4%;花芽坐果率比不间伐改形提高23.4百分点;果实单果质量、硬度分别比不间伐改形分别提高16.7%、17.3%;着色面积和可溶性固形物分别比不间伐改形提高12.3,1.03百分点。间伐改形后花芽在冠层中的立体空间分布均衡,有效改善了不间伐改形树体花芽外移和上移现象。可见,间伐改形不仅能够有效解决密闭果园枝量繁多、树形紊乱等问题,还可有效改善枝条组成比例,提高花芽质量及坐果率,明显提高果实品质。     

低温解除牡丹休眠进程中基因组DNA甲基化敏感扩增多态性(MSAP)分析

DNA甲基化是表观遗传调控的重要组成部分,在真核生物的基因表达调控中发挥了重要作用。本实验研究了感受0、6、12、18和24d4℃低温的牡丹(Paeonia suffruticosa)鲁荷红移入温室后的萌动和开花表现,并对其进行了甲基化敏感扩增多态性(methylation sensitive amplified polymorphism,MSAP)分析。结果表明,4℃低温处理18d可解除鲁荷红内休眠,继续增加低温则进入生态休眠阶段。MASP分析表明,牡丹内休眠期间花芽内DNA甲基化程度很高,甲基化位点占总位点比率的60%以上,以半甲基化为主,在低温处理进程中,甲基化水平总体呈下降趋势。与未经低温的对照相比,约50%的位点发生了甲基化模式变化,低温6、12、18和24d去甲基化位点数和比例分别为97(17.1%)、104(18.6%)、148(24.6%)和218(36.1%),过甲基化的位点数分别为197(34.7%)、137(24.6%)、95(15.8%)和79(13.1%)。研究结果提示,DNA甲基化参与了牡丹内休眠的调控。     

甘蓝花粉特异表达的多聚半乳糖醛酸酶基因BoMF9的克隆与特征分析

根据白菜花粉特异的多聚半乳糖醛酸酶基因BcMF9的全长序列设计引物，通过PCR直接扩增的方法从结球甘蓝（Brassica. oleracea var. capitata）和芥蓝（B. oleracea var. alboglabra）中克隆得到BcMF9的同源基因BoMF9（BoMF9c和BoMF9l）。生物信息学分析和系统树构建发现BoMF9c和BoMF9l具有花粉特异表达的多聚半乳糖醛酸酶基因的序列特征。不同组织的表达特征分析发现它们分别在结球甘蓝和芥蓝的花蕾中特异表达。推测BoMF9c和BoMF9l可能在花粉萌发和花粉管伸长中起作用。     

紫菜薹快速碱化因子基因BcMF14p的序列与表达分析

快速碱化因子是近年来新发现的一种植物多肽类信号分子。根据已知的普通白菜两用系中可育株系特异表达的快速碱化因子基因BcMF14序列,在其编码框两侧设计引物,从紫菜薹中克隆出该类信号分子基因,命名为BcMF14p(登陆号:EF523517),全长273 bp,不含内含子,编码框包含222个核苷酸,编码73个氨基酸。NCBI在线同源比对结果表明该基因属于快速碱化因子家族,与花椰菜、拟南芥等的快速碱化因子核苷酸序列相似性达到86%以上。对该信号分子的表达进行RT-PCR分析,结果显示BcMF14p只在紫菜薹花蕾、花、角果等生殖器管中表达,在茎、叶等营养器官中不表达,说明BcMF14p可能参与紫菜薹生殖发育过程中的信号传导。     

Structural requirements of Angiotensin I-converting enzyme inhibitory peptides: quantitative structure-activity relationship study of di- and tripeptides

A database consisting of 168 dipeptides and 140 tripeptides was constructed from published literature to study the quantitative structure--activity relationships of angiotensin I-converting enzyme (ACE) inhibitory peptides. Two models were computed using partial least squares regression based on the three z-scores of 20 coded amino acids and further validated by cross-validation and permutation tests. The two-component model could explain 73.2% of the Y-variance (inhibitor concentration that reduced enzyme activity by 50%, IC50) with the predictive ability of 71.1% for dipeptides, while the single-component model could explain 47.1% of the Y-variance with the predictive ability of 43.3% for tripeptides. Amino acid residues with bulky side chains as well as hydrophobic side chains were preferred for dipeptides. For tripeptides, the most favorable residues for the carboxyl terminus were aromatic amino acids, while positively charged amino acids were preferred for the middle position, and hydrophobic amino acids were preferred for the amino terminus. According to the models, the IC50 values of seven new peptides with matchable primary sequences within pea protein, bovine milk protein, and soybean were predicted. The predicted peptides were synthesized, and their IC50 values were validated through laboratory determination of inhibition of ACE activity.     

芥蓝BaCSLD的克隆、亚细胞定位及表达分析

为获知芥蓝中类纤维素合成酶基因的序列特征和表达特性,利用RT-PCR技术从芥蓝中扩增获得1个类纤维素合成酶基因全长cDNA序列,命名为BaCSLD (GenBank 登录号:MH491418),对该基因的序列特征、亚细胞定位、组织特异性表达情况进行分析,并通过最大似然法构建系统进化树。结果表明,BaCSLD cDNA序列包含一个2 796 bp的最大开放阅读框,编码931个氨基酸,且BaCSLD属于跨膜蛋白,具有8个跨膜区。BaCSLD与12个物种同源序列在进化树上分为4组,来自十字花科的AtCSLD与BaCSLD处于同一分支,亲缘关系较近的为苦瓜中的McCSLD,而与同属的油菜中的BnCSLD和芜菁中的BrCSLD关系较远。BaCSLD::GFP融合蛋白定位于细胞膜。此外,BaCSLD仅在芥蓝三级花蕾和花中表达。本研究结果为类纤维素合成酶基因功能的深入研究提供了参考。     

贵州红枫湖溶解有机质不同分离组分的卤代活性研究

饮用水使用氯化消毒产生的消毒副产物给人类健康带来了极大危害。利用XAD树脂与阴、阳离子交换树脂对贵州红枫湖水体中溶解有机质进行了富集分离,分成腐植酸、富里酸、疏水中性物质及亲水酸性、碱性及中性物质6种组分,比较了各组分在加氯消毒过程中卤代活性的大小及其三卤甲烷的生成情况,揭示了有机组分卤代活性与其结构之间的关系,并探讨了它们可能的来源,提出了相应的控制措施。结果表明,红枫湖水体溶解有机质以富里酸组分为主,占分离出总有机碳的56%。氯化消毒实验发现消毒副产物以三氯甲烷和一溴二氯甲烷为主,富里酸的卤代活性最强。结构分析显示各有机组分的卤代活性与紫外吸光系数和酚羟基含量之间具有显著相关性。6种分离组分的消毒副产物生成量中,富里酸是生成三卤甲烷的主要前驱物质,占消毒副产物总量的76%。有机组分的碳稳定同位素值与C/N比值表明,富里酸主要来自陆源有机物,其余组分兼有陆源和内源两种来源。因此,控制陆源污染是减少红枫湖消毒副产物前驱物的有效途径。     

Effects Of Potassium,Phosphorus And Nitrogen Fertilization On Endogenous Ethylene And Quality Characteristics Of Apples (Malus Domestica L.)

Factor analysis of the data from experiment one with fertilization of ‘Spartan’ apples using various levels of nitrogen, phosphorus, and potassium and their interactions showed significances between ethylene and potassium, soluble solids and potassium, acidity and phosphorus, ethylene and phosphorus, and between ethylene and nitrogen. Factor analysis of data from experiment two with twenty apple cultivars showed that maturation time and ethylene were positively associated, whereas ethylene was inversely related to acidity that is reduced during the apple fruit maturation, and soluble solids are produced during apple fruit maturation. The effects of potassium on ethylene could be through promotion of enzyme activation, promotion of protein synthesis and increased solute transport in the xylem and phloem. Production of ethylene through soluble solids could be due to increased synthesis of the amino acid cysteine that is precursor of ethylene that may be extended to synthesis of other amino acids. The decreasing concentration of malic acid during fruit maturation may be due to increasing rates of several enzymes involved in the citric acid and glyoxylate cycles that are delivering precursors to synthesis of amino acids and porphyrins. The significant relationships between soluble solids and potassium and between acidity and phosphorus may increase the synthesis of nucleic acids and improve energy supply through adenosine phosphates. The significance between ethylene and nitrogen may also increase synthesis of amino acids, proteins and enzymes. On this basis it may be concluded that the three major minerals nitrogen, phosphorus and potassium are very important for apple fruit maturation because they may be involved in several biochemical processes.     

The role of root exudates in specific apple (Malus × domestica Borkh.) replant disease (SARD)

In an effort to improve our understanding of the specific apple replant disease (SARD), direct and indirect effects of phytohormones and related compounds (abscisic acid, 6‐benzyladenine, indole‐3‐acetic acid, 1‐naphthaleneacetic acid, and gibberellins GA₃ or GA₄) on root exudates of apple seedlings were evaluated as a potential mediating factor in the infection process. In the first type of experiments, radioactively labeled hormonal substances were applied to the stumps of decapitated apple‐seedlings and the occurrence of radioactivity in root exudates and their qualitative characterization were examined (direct influence). In another set of experiments, the effects of leaf‐applied plant growth regulators on the amount and composition of the predominant organic acids, carbohydrates, and amino acids/amides were studied. Cherry seedlings resistant to SARD and apple seedlings with dormant apical buds and, thus, not susceptible to infection were used for comparison. The results showed no differences in exudation of applied plant growth regulators between growing cherry and apple seedlings. Thus, a direct effect of plant hormones on the infection process is unlikely. However, leaf treatments with growth regulators, in particular with auxin‐type compounds and abscisic acid, increased exudation of alditols. This may indicate that plant hormones are, to some degree, indirectly involved in the infection process.     

How potassium deficiency alters flower bud retention on cotton (Gossypium hirsutum L.)

Potassium (K) is fundamental for plant growth and development but despite the increased quantities of fertilizers applied, incidents of K deficiency are commonly observed. The objective of this study was to record the effects of K deficiency during cotton’s (Gossypium hirsutum L.) early reproductive stage on carbohydrate content and metabolism, total antioxidant capacity and oxidative damage of cotton flower buds as well as the physiology of the leaf, subtending to the flower buds. Growth chamber experiments were conducted using cotton cultivar DP0912 and treatments consisted of normal K and deficient K fertilization for the duration of the experiment. Potassium deficiency resulted in significant oxidative damage in the cotton flower buds, despite the substantial increase in their total antioxidant capacity. Sucrose metabolism of the flower buds was markedly affected resulting in significant reductions in all non-structural carbohydrate concentrations. Furthermore, K deficiency disturbed leaf physiology leading to increased membrane damage, decreases in chlorophyll and carotenoid content and ultimately leaf photosynthetic rates. Concomitant increases in specific leaf weight under K deficient conditions indicated reductions in photoassimilate translocation, which in conjunction with the disruptions observed in flower bud sucrose metabolism, due to the insufficient antioxidant response, resulted in significant decreases in flower bud retention.     

Corn protein-based thermoplastic resins: effect of some polar and amphiphilic plasticizers.

Homogeneous blends of corn gluten meal (CGM) and "polar" plasticizers (water, glycerol) or "amphiphilic" plasticizers [octanoic and palmitic acids, dibutyl tartrate and phthalate, and diacetyl tartaric acid ester of mono-diglycerides (DATEM)] were obtained by a hot-mixing procedure. The glass transition temperature (T(g)) of the blends was measured by modulated differential scanning calorimetry and dynamic mechanical thermal analysis, as a function of plasticizer type and content (0-30%, dwb). The plasticizing efficiency (i.e., decrease of T(g)) at equal molar content was found to be proportional to the molecular weight and inversely proportional to the percent of hydrophilic groups of the plasticizer. The migration rate of the plasticizers in the polymer was related to their physicochemical characteristics. It was assumed that polar substances interacted with readily accessible polar amino acids, whereas amphiphilic ones interacted with nonpolar zones, which are buried and accessible with difficulty. The temperature at which a thermoplastic resin of plasticized CGM could be formed was closely connected to the T(g) of the blend.     

不同品种花生油脂体粒径电位和蛋白质组成的分析

为了探索不同品种花生油脂体的物理和化学性质差异,以5种（豫花23,豫花27,豫花9719,豫花9830和豫花9502）油脂含量不同的花生品种为原料,采用水剂法提取油脂体,并对提取后油脂体的粒径、ζ电位、氨基酸组成、蛋白质分子量分布进行分析比较。结果表明：提取后,5种花生油脂体粒径间存在一定差异,以豫花9719的粒径较大;花生油脂体均含有油脂体蛋白和贮藏蛋白,但不同品种间存在蛋白质种类的差异;5种花生油脂体在pH值为3.0时ζ电位为正值,在pH值为7.4和pH值为9.0时ζ电位为负值,盐浓度的增加会降低油脂体ζ电位的绝对值;5种花生油脂体的蛋白质均为极性带负电氨基酸质量分数均大于非极性带正电或不带电氨基酸,但氨基酸总量各不相同,以豫花27较低。该研究可为花生油脂体的品质特性研究和应用产品开发提供参考。     

Differences in the chemical composition of dissolved organic matter from waste material of different sources

The chemical composition of waste-material-derived dissolved organic matter (DOM) was characterized by chemolytic analyses and ¹H, ¹³C and ³¹P nuclear magnetic resonance (NMR) spectroscopy. Dissolved organic matter was extracted by water from an aerobic fermented urban waste compost, a sewage sludge and a pig slurry and then fractionated using the XAD-8 method. The amount of water-extractable dissolved organic carbon (DOC) ranged from 3% in the sewage sludge to 22% in the pig slurry. Dissolved organic matter isolated from pig slurry was equally distributed between hydrophilic and hydrophobic DOC, whereas in the sewage-sludge-derived material the hydrophobic fraction was predominant. Dissolved organic C from the urban waste compost was mainly within the hydrophilic fraction. Wet-chemical analysis and ¹H- and ¹³C-NMR spectra showed that both DOM fractions from the urban waste compost were low in neutral, acidic and amino sugars as well as in lignin-derived compounds. In turn, the materials were rich in low-molecular-weight aliphatic compounds. The chemical structure of both fractions is probably the result of the intensive transformation of urban waste compost during its fermentation. The hydrophilic fractions of DOM from sewage sludge and pig slurry contained considerable amounts of carbohydrates but were also rich in low-molecular-weight aliphatics. The respective hydrophobic fractions had the largest contents of CuO-extractable phenols which may in part derive from sources other than lignin. By contrast with the other materials, the hydrophobic fraction from the pig slurry seemed to contain polymeric rather than low-molecular-weight material. The ³¹P-NMR spectrum of the hydrophilic DOM fraction from urban waste compost did not show signals of inorganic or organic P compounds while the spectrum of the hydrophobic fraction revealed traces of monoester P, diester P, and orthophosphate. ³¹P-NMR spectroscopy suggested that both the hydrophobic and hydrophilic fractions from pig slurry did not contain organic P. The hydrophilic DOM fraction from sewage sludge contained orthophosphate, organic monoester P and a little pyrophosphate. The hydrophobic fraction contained mainly organic diester P and smaller amounts of teichoic acids and organic monoester P. Considering that water-soluble fractions of urban waste compost contained no easily plant-available P and a low content of labile organics, we conclude that this material contains less labile nutrients and is more refractory than the soluble constituents of pig slurry and sewage sludge.