银杏营养贮藏蛋白质的分离鉴定

采用光学和电子显微镜技术、SDS-PAGE技术、间接免疫酶标光镜定位和胶体金免疫电镜定位技术对银杏枝条中的营养贮藏蛋白质(VSPs)进行分离鉴定,并对其细胞学特性及动态变化规律进行初步研究。研究结果:运用制备型SDS-聚丙烯酰胺凝胶电泳分离并制备银杏32和36ku2种营养贮藏蛋白质抗原,通过免疫新西兰大白兔成功获得银杏32和36ku2种营养贮藏蛋白质的多克隆抗体。银杏VSPs主要分布于落叶期当年生枝条的次生韧皮部细胞中,呈现出团块状、颗粒状和均一状3种形态。在新梢生长期,积累在枝条中的VSPs逐渐降解消失,表现出明显的季节变化。     

银杏营养贮藏蛋白质的细胞学研究

采用透射电子显微技术,对银杏的末端小枝和根中贮藏蛋白质细胞进行了超微结构的观察,研究结果表明:银杏营养贮藏蛋白质主要在枝条和根次生韧皮部的韧皮薄壁细胞和韧皮射线细胞中积累;液泡中的蛋白质主要以絮状、不规则块状、颗粒状和均一细沙状的形式存在;落叶期,银杏枝条贮藏蛋白质大量积累。2月底3月初,随着芽的萌动和新梢的生长,枝条中贮藏蛋白质首先被动用,根中的贮藏蛋白质积累和动用都晚于枝条。5月份,新梢中也开始积累贮藏蛋白质;内质网、高尔基体、线粒体与贮藏蛋白质的降解过程有关。     

银杏枝条营养贮藏蛋白质的免疫细胞化学鉴定

依据银杏枝条中32和36 ku蛋白质含量的明显季节变化,推测它们是银杏的营养贮藏蛋白质.采用间接免疫组织细胞化学定位技术,对这2种蛋白质进行细胞化学定位.32和36 ku蛋白质具有一定的免疫相关性,它们分布在一些皮层薄壁细胞和韧皮部的韧皮薄壁细胞中,存在于这些细胞的小液泡里.研究结果揭示这2种蛋白质的组织分布式样,为这2种蛋白质是银杏的营养贮藏蛋白质提供有力证据.     

印度紫檀营养贮藏蛋白质的主要特性

采用光学和电子显微镜技术、十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术分离鉴定热带落叶树木印度紫檀营养贮藏蛋白质(VSPs),并采用免疫印迹技术分析包括印度紫檀在内的5种热带树木VSPs之间的免疫相关性.结果表明:印度紫檀VSPs包括4种蛋白质,分子质量分别为33.5、28、21和19.5 ku;它们主要分布在枝条木薄壁细胞、木射线薄壁细胞、韧皮薄壁细胞和韧皮射线薄壁细胞、皮层薄壁细胞以及树干、大根的韧皮薄壁细胞和韧皮射线薄壁细胞中;它们的超微结构存在明显差异,大致可以分为电子致密的团块状、絮状和均一状3种形态,不同形态的VSPs存在于不同的贮藏蛋白质细胞中,具有细胞特异性;在新梢发育时期,积累在枝条中的VSPs明显消失,但积累在树干和大根中的VSPs和落叶期相比没有明显变化,积累在枝条中的营养贮藏蛋白质是新梢生长发育所需氮素的主要来源;同一科树木的营养贮藏蛋白质之间存在较强的免疫相关性,但不同科树木的营养贮藏蛋白质之间缺乏免疫相关性.树木营养贮藏蛋白质生化性质的多样性可能反映了生物学功能的多样性.     

旱柳枝条叶绿体光化学特征的径向异质性

【目的】研究枝条叶绿体光化学特征的径向异质性和组织特异性,可为确定枝条中不同组织对枝条碳同化的贡献以及揭示叶绿体对枝条异质性光环境的适应机制提供依据。【方法】测定照光和黑暗条件下旱柳无性系当年生枝条的CO_(2 )释放速率,评价枝条光合的碳回收贡献。测定树皮绿色组织、木质部和髓心的光合色素含量、吸光系数、光化学效率、电子传递速率,分析不同组织中叶绿体光化学特征的差异。【结果】饱和光强下旱柳当年生枝条的总光合速率达到1.27μmolCO_2·m^-2s^-1,可将77%呼吸消耗的碳回收固定。在旱柳当年生枝条中,树皮的总叶绿素含量显著高于木质部和髓心,分别是木质部和髓心的15.90和1.83倍。叶绿素b与叶绿素a的比例随径向深度的增加由树皮至髓心呈显著升高的趋势,树皮中的类胡萝卜素含量、类胡萝卜素与叶绿素的比例均显著高于木质部。旱柳当年生枝条不同组织的吸光系数沿径向方向显著降低,树皮绿色组织的最大光化学效率、实际光化学效率和相对电子传递速率显著高于木质部和髓心。【结论】枝条光合可将77%呼吸消耗的碳回收。不同组织的光合色素含量和光化学效率呈现随径向深度增加而显著降低的规律,树皮绿色组织是枝条光合的主要载体。树皮绿色组织具有较强的光保护能力,木质部和髓心通过光合色素比例的调整最大限度地捕捉组织中有限的光能。     

兴安落叶松不同器官中的黄酮含量的动态变化

黄酮是植物体内一类重要的次生代谢产物,具有重要的生态作用和经济价值.但目前对兴安落叶松黄酮的研究很少.本论文研究了自然条件下兴安落叶松黄酮动态变化的特征,并对黄酮与温度和光照的相关关系进行初步探讨.对兴安落叶松不同器官黄酮含量的分析结果表明:(1)树皮,茎木质部,针叶和枝中黄酮含量的高低顺序为:树皮(7.78%) >针叶(2.79%)>枝条(1.72%)>茎木质部 (1.19%).(2)树皮,茎木质部,针叶和枝中黄酮含量具有明显的季节变化,但各器官(树皮,树叶,树枝,茎木质部)黄酮含量对温度的依赖性并不相同.就树皮而言,随着温度的升高,黄酮的含量与温度呈现较明显的正相关关系(R2=0.75).但其它器官黄酮含量对温度的依赖性不强;(3).随着季节的变化,黄酮含量的最高值出现在夏秋季节,含量是冬春季节的3-4倍.这种现象可能归因于夏季落叶松遭受的多种环境胁迫,例如紫外辐射,高温等可能促进了黄酮的积累.(4)不同高度和方向的针叶中,黄酮含量阳生叶比阴生叶多,上部叶比下部叶多,说明阳光辐射促进了黄酮的积累(R2=0.76).总而言之黄酮在落叶松抵御自然环境的胁迫中发挥了积极的作用,保护落叶松免受高温和强光的伤害,但是不同器官的黄酮可能发挥着不同的作用.     

不同梓属树种花器官蛋白电泳分析

为了分析与楸树自交不亲和有关的特异蛋白质特性,采用考马斯亮蓝染色法和SDS-PAGE技术研究了2种梓属树种(滇楸、楸树)花器官的蛋白质含量及蛋白质组分。结果表明:楸树-1(连云港种源)的花柱、子房、花萼蛋白质含量较低,滇楸和楸树-2(老山种源)较高,而楸树-1花瓣蛋白质含量略高于其他树种;同一树种不同器官蛋白质含量表现为子房>花萼>花柱>花瓣。蛋白组分分析表明,梓属各树种不同花器官内的蛋白质主要集中在12～45 ku之间,各器官共有蛋白20个左右,发现具有特异性的蛋白包括:楸树-1花柱42 ku蛋白,楸树-2花柱64和19 ku蛋白,滇楸花柱37 ku蛋白,以及楸树-2和滇楸花柱共有的58和24 ku蛋白;楸树-1子房41、38和23 ku蛋白;花萼中,88和65 ku蛋白为楸树-1所有,52、40、26 ku蛋白为楸树-2和滇楸特有。花瓣内蛋白带较少,仅发现63 ku蛋白为楸树-1特异蛋白。     

银杏枝条部位和年龄对不定根形成的影响及其与非结构碳水化合物含量的关系

[目的]研究银杏枝条部位、年龄对不定根形成的影响,并探讨其相应的叶、茎段中非结构碳水化合物(NSC)含量变化与扦插生根的关系。[方法]以50年生银杏树冠上、下部枝条的1、2和3年生位置的茎段为材料截取插穗,500 mg·L-1IBA与纯水对照处理基部60 s,在全光喷雾状态下研究扦插生根情况,并利用HLPC法分析相应部位叶和茎段中NSC含量。[结果]表明:银杏扦插不定根形成过程分为4个阶段,即0～4周愈伤形成期、4～6周愈伤膨大期、6～8周不定根开始形成期、8周后不定根伸长;生根部位主要在切口处。树冠上、下部位插穗的生根率未见明显差异,但随着枝龄的增加而明显降低。NSC分析表明:下部叶片NSC组分中多糖和总糖含量明显高于上部的,但茎段NSC的所有组分在上下部均无明显差异;随着枝龄的增加,相应位置叶片NSC各组分含量没有明显差异,但相应茎段NSC组分中葡萄糖、果糖、多糖、总糖、淀粉含量却明显降低,与生根率的变化趋势一致。此外,500 mg·L-1IBA处理并未明显提高生根率,还需进一步优化处理浓度。[结论]银杏树冠上部和下部枝条对不定根形成无影响;未处理枝条的枝龄与生根率呈负相关,而茎段NSC含量与生根率呈正相关;叶片NSC含量对不定根形成影响较小;生根方式以愈伤生根类型为主。该结论将为今后银杏扦插生根及其机制研究提供参考。     

不同时期修剪对早实核桃枝条生长特性及其营养物质含量的影响

为确定早实核桃最佳修剪时期,保证连年丰产、稳产,以河北绿岭果业有限公司长势健壮的6a生‘绿岭’核桃树为试材,研究了不同时期修剪对‘绿岭’核桃枝条生长特性及其可溶性糖、可溶性蛋白含量的影响,结果表明:秋季修剪后枝条萌发结果母枝的数量高于春季萌芽前修剪,但效果不显著;春季萌芽前修剪后萌发结果母枝的平均粗度为2.31cm,形成混合花芽的平均数量为201个,二者均高于秋季修剪;春季萌芽前修剪后枝条剪口的平均风干高度为0.36cm,显著低于秋季修剪;春季萌芽前修剪后2.5~3.5cm粗度枝条皮层可溶性糖含量和1.5~2.5cm粗度枝条木质部可溶性糖含量分别为6.13%、3.81%,均显著高于秋季修剪;春季萌芽前修剪后,除5.5~6.5cm粗度枝条外,其他4个粗度枝条木质部的可溶性蛋白含量分别为0.15%、0.30%、0.17%、0.26%,均显著高于秋季修剪。由此可知,春季萌芽前修剪能促进结果母枝的增粗生长和混合花芽的形成,降低修剪口风干长度,利于枝条中可溶性蛋白含量及1a生枝中可溶性糖含量的积累,因此,‘绿岭’核桃适宜修剪时期为春季萌芽前半个月。     

树干注药后吡虫啉在枣树不同部位的传导分布动态

为了阐明吡虫啉在金丝小枣树体内的传导机理,在树干滴注林果注干杀虫剂后,测定其不同方位叶片、枝条木质部及韧皮部中的吡虫啉含量。结果表明,经树干滴注杀虫剂后,吡虫啉在枣树叶片、枝条木质部及韧皮部组织中均具有良好的传导、分布性能,随着时间的延长,枣树叶片、枝条木质部及韧皮部中的吡虫啉含量均表现出先上升后下降的变化趋势;注药后21 d、15 d、15 d枣树叶片、枝条木质部及韧皮部中的吡虫啉含量分别达到最高值0.90 mg/kg、1.03 mg/kg、0.83 mg/kg。然后逐渐降低,均于用药70 d后变化趋于平缓,而注药后85 d时,枣树叶片、枝条木质部及韧皮部中的吡虫啉含量分别为0.79 mg/kg、0.54 mg/kg、0.64 mg/kg,仍处于较高水平;枣树不同部位吡虫啉平均含量由大到小依次为叶片、枝条木质部、枝条韧皮部,叶片中的吡虫啉平均含量为0.70 mg/kg,比枝条木质部和韧皮部分别增加25.00%和45.83%,木质部平均含量为0.56 mg/kg,比韧皮部含量增加16.67%;树冠上层叶片、上层枝条木质部及上层枝条韧皮部中的吡虫啉平均含量分别达到0.78 mg/kg、0.58 mg/kg、0.49 mg/kg,均高于下层的含量,说明吡虫啉在树体内传输时纵向传导能力较强。枣树经树干滴注吡虫啉后,药效持续时间长,可有效防治枣树生长期内的主要害虫。     

红豆杉枝叶中产物分布研究

研究了南方红豆杉枝叶中叶片、树皮、嫩枝和木质部的生物量比例,以及各部位中主要有效成分紫杉醇和10-DABⅢ的含量分布情况。实验结果表明:红豆杉枝叶中叶片的生物量约占30%,叶片中累积的有效产物量最高(约占总量的70%);树皮含量次之,杂质最少;红豆杉枝叶中木质部的生物量最多(占46%),累积的产物最少(约占总量的5%)。叶片是红豆杉枝叶中提取紫杉醇等产物的最有效部位。     

Distribution and partial characterization of seasonally expressed proteins in different aged shoots and roots of 'Loring' peach (Prunus persica)

During autumnal leaf senescence, leaf nitrogen in deciduous trees is translocated to storage sites, especially bark and xylem tissues. Proteins that accumulate in large amounts in bark and xylem in winter and are absent from these organs in summer are called storage proteins, and are believed to be vehicles for storing nitrogen reserves. These reserves are important for spring growth and help trees tolerate or recover from both abiotic and biotic stresses. Based on seasonal patterns of accumulation, we previously identified three storage proteins with molecular masses of 60, 19 and 16 kDa in bark tissues of 'Loring' peach (Prunus persica (L.) Batsch). To characterize the distribution of these proteins in different-aged tissues and to determine if they have any function other than nitrogen storage, we examined their seasonal distribution in bark tissues of current-year and 1-year-old shoots, scaffold branches, main trunks and 4-5-year-old roots of 'Loring' peach. Verification of protein identity was based on molecular mass and reactions with antibodies directed against each specific protein. Protein distribution was variable. For all three proteins, the greatest amount was present in mid-winter in current-year and 1-year-old shoots. These tissues also showed the greatest seasonal variation in the amount of protein present. The 16 kDa protein was present only in the youngest shoots, whereas the 19 kDa protein was present in all tissues examined. The 60 kDa protein was absent in root tissue. The amino acid composition and sequence of each protein were determined. The 60 kDa protein was identified as a dehydrin, and the 19 kDa protein appeared to be related to a family of allergen proteins in Rosaceous plants, some members of which are associated with pathogenesis-related proteins. The amino acid sequence of the 16 kDa protein appeared to have no homology with any proteins in the SwissProt database. Therefore, it is likely that the 16 kDa protein, in a strict sense, is a bark storage protein. Defining storage proteins solely by their pattern of accumulation and the extent to which they accumulate may not be a good functional definition. It is possible that storage proteins have functional roles in addition to nitrogen storage.     

杉木人工林各植物组分含碳率研究

分别采集会同县广坪林区第2代杉木人工林的杉木树干(去皮)、树皮、枝、叶、根系样品和灌木的干、枝(叶)和根样品与草本植物的地上、地下部分样品以及林内的枝、叶、果和碎屑凋落物和枯死根,用干烧法测定其含碳率。结果表明:杉木各器官含碳率大小的顺序是:树皮>树叶>树根>树干>树枝。树皮、树叶、树干、树枝含碳率随着树木年龄的增大而增加,树根含碳率随着树木年龄增大出现波动。林冠下草本植物的平均碳含量比木本植物低,且草本植物间碳含量差异要比木本植物间大。林龄对凋落物同种组分碳含量影响不显著。不同凋落物的碳含量即使在同一龄级也存在较大差异。枯死根系的碳含量要低于地上凋落物各组分的碳含量,林木各器官活有机体内的碳含量均大于相应死有机体(凋落物)内的碳含量。     

A potential role for xylem-phloem interactions in the hydraulic architecture of trees: effects of phloem girdling on xylem hydraulic conductance

We investigated phloem-xylem interactions in Acer rubrum L. and Acer saccharum Marsh. Our experimental method allowed us to determine xylem conductance of an intact branch by measuring the flow rate of water supplied at two delivery pressures to the cut end of a small side branch. We found that removal of bark tissue (phloem girdling) upstream of the point at which deionized water was delivered to the branch resulted in a decrease (24% for A. rubrum and 15% for A. saccharum) in branch xylem hydraulic conductance. Declines in hydraulic conductance with girdling were accompanied by a decrease in the osmotic concentration of xylem sap. The decrease in xylem sap concentration following phloem girdling suggests that ion redistribution from the phloem was responsible for the observed decline in hydraulic conductance. When the same measurements were made on branches perfused with KCl solution (approximately 140 mOsm kg(-1)), phloem girdling had no effect on xylem hydraulic conductance. These results suggest a functional link between phloem and xylem hydraulic systems that is mediated by changes in the ionic content of the cell sap.     

兴安落叶松单宁含量的器官差异与季节变化

兴安落叶松是生产落叶松单宁的重要原材料,随着天保工程的实施,使得主要原料树皮供应紧张。通过2003～2004年的测定结果表明,除现用的树皮(8.8%～15.4%)生产原料外,根系(约7.8%)、树枝(1.0%～7.0%)甚至叶片(4.0%～8.0%)都可以成为生产原料,而树干木质部含量过低(0.6%～1.4%)不宜成为生产原材料。不同季节单宁含量差异结果表明,秋季(9～10月)的落叶松单宁含量都显著高于其它季节,因此,原材料的收集应该在秋季。此外,叶片单宁含量与光照强度呈现显著正相关。     

Influence of stem rot pathogen <Emphasis Type="Italic">Fomitiporia</Emphasis> sp. on “Sanbu-sugi” cultivar of the Japanese cedar <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

An unidentified Fomitiporia sp. causes severe white-rot on stems of a cultivar “Sanbu-sugi” of the Japanese cedar, Cryptomeria japonica. The influence of the fungus on tree health and wood properties was determined. Bark from dead branches and xylem from living branches contained more glucose than bark from living branches and xylem from dead branches. Tree heights at which annual rings were disconnected were 2, 4 and 6 m at ages 37, 15 and 24 years old, respectively. The pH values of damaged parts were lower than those of non-damaged parts, and the damaged parts were clearly identified using bromocresol green solution. Weight loss of sapwood during 60 days of fungal degradation was 1.4 %, which was less than that by a saprophyte, Trametes versicolor. The amount of polyphenols in the heartwood from damaged tree stems was less than that from non-damaged stems. Degraded parts were less stiff than the non-degraded sapwood as measured with a wood-decay tester, Pilodyn. Our observations indicate that damaged stems are chemically and physically inferior to non-damaged stems.     

Multiscale analysis of the contents of palmatine in the Nature populations of Phellodendron amurense in Northeast China

Palmatine is a valuable ingredient in Chinese medicine that is produced by Phellodendron amurense Rupr.The contents of palmatine content in root bark, trunk bark,perennial branch bark, annual branches, and leaves of the trees with different ages and geographies in Northeast China were measured by high-performance liquid chromatography.The contents of palmatine in the barks of root, trunk, and perennial branch were significantly higher than those in annual branches and leaves. The contents of palmatine in trunk bark and root bark from Lesser Khingan Mountains increased with age, which is significantly opposite to other three vegetation types. The contents of palmatine in perennial branch bark, annual bark and leaves had no significant regularity. Moreover, the contents of palmatine in samples of root bark, trunk bark, perennial branch bark and annual bark varied significantly with latitude. The nature populations of P. amurense growing at low latitude contained significantly more palmatine than those growing at high latitude. These results provide a scientific basis for the reasonable cultivation and efficient utilization of P. amurense.