NaCl对大麦幼苗根系蛋白质和游离氨基酸含量的影响

大麦幼苗在２００ｍｍｏｌ／ＬＮａＣｌ处理６ｄ过程中,根中蛋白水麦活性下降,可溶性蛋白含量在处理１、２、４ｄ下降,６ｄ时有所提高。多数游离氨基酸相对含量及其总量呈上升势,胁迫１、２、６ｄ时,总量较对照分别上升６４．２０％、５６．６９％和１．６９％,其中Ｐｒｏ、Ｇｌｕ、Ａｌａ和Ｔｈｒ较为突出,盐胁迫下根系质膜和液泡膜结合慢白含量上升。可溶性蛋经ＳＤＳ－ＰＡＧＥ电泳和扫描分析,在盐胁迫１、２、６ｄ中２５     

水分胁迫对小麦叶绿体色素蛋白复合体的影响

水分胁迫可抑制激发能向ＰＳⅡ传递,降低ＰＳⅡ捕光色素蛋白复合体、ＰＳⅡ内周天线色素蛋白复合体以及ＰＳⅠ捕光色素蛋白复合体和ＰＳⅠ反应中心叶绿素ａ 蛋白复合体的含量,其中以ＰＳⅡ色素蛋白复合体含量下降幅度较大。类囊体膜多肽分析表明,水分胁迫使属于ＰＳⅡ捕光色素蛋白复合体的２５ ｋＤ蛋白、ＰＳⅡ内周天线色素蛋白复合体的４３ ｋＤ 和４７ ｋＤ蛋白以及ＰＳⅠ捕光色素蛋白复合体的２１ ｋＤ蛋白含量降低,尤以２５ ｋＤ蛋白含量降幅最大。     

磷饥饿下番茄幼苗根系液泡膜H+-ATPase活性的适应性变化

以‘世纪星’番茄为材料。研究了磷饥饿下番茄幼苗的生长状况及其根部液泡膜H^＋-ATPase活性的适应性变化。结果表明：磷饥饿下番茄幼苗的平均高度均低于对照苗,而主根长度均显著长于对照。磷饥饿提高了番茄幼苗根部液泡膜H^＋-ATPase的水解活性,随着磷胁迫时间的延长,该酶的活性逐渐增大,在磷饥饿7d时达到最大,后又略有降低;而对照番茄幼苗根部该酶的活性变化很小。动力学分析表明：磷饥饿使番茄幼苗根部液泡膜H^＋-ATPase的Km值明显降低,但对该酶的Kmax影响不大。这说明磷饥饿提高了该酶对其底物的亲和力。此外,磷饥饿并不改变液泡膜H^＋-ATPase酶的最适pH值（仍为7．5）。     

磷饥饿时番茄幼苗根系质膜H^＋—ATP酶活性的变化与磷吸收的关系

磷饥饿提高了番茄幼苗质膜H^ -ATP酶活性并促进了番茄幼苗根部的H^＋分泌和。动力学分析表明,磷饥饿使番茄功苗根部质膜H^ -ATP酶的K m值明显降低,亦即提高了该酶对其底物的亲和力,但对该酶的Vmax影响不大。另外,磷饥饿并不改变ATP酶的最适pH值（最适pH值为6.5）。钡酸盐显著抑制番茄幼苗根部质膜ATP酶的活性以及H^＋分泌,也显著抑制番茄幼苗的Pi吸收。与对照相比,上述抑制作用在饥饿处理的植物中表现得更强,以上结果表明,磷饥饿时高亲和性Pi传递系统的诱导很可能包含质膜H^ -ATP酶的参与。     

增强UV—B辐射对菜豆蛋白质代谢的影响

模拟研究了温室条件下兰州地区臭氧层减薄２５ ％ 时增强的ＵＶＢ（２８０ ～３２０ ｎｍ） 辐射对菜豆（ Ｐｈａｓｅｏｌｕｓｖｕｌｇａｒｉｓ Ｌ．） 蛋白质代谢的影响。菜豆幼苗期,ＵＶＢ辐射（ＵＶＢＢＥ：１６ ｋＪ·ｍ －２·ｄ－１） 促进菜豆叶片中蛋白质的合成;菜豆生长后期,ＵＶＢ辐射则抑制蛋白质的合成,降低叶片中可溶性蛋白质的含量,促进蛋白水解酶活性的上升和总游离氨基酸的积累。ＳＤＳＰＡＧＥ分析表明,ＵＶＢ辐射使幼苗期菜豆叶片中８８ ｋＤ、７６ ｋＤ、４２ ｋＤ、３５ ｋＤ、３３ ｋＤ 多肽的含量上升;而在菜豆生长后期,ＵＶＢ辐射使大分子量多肽（７６ ｋＤ 以上） 的含量迅速下降,１０～１４ ｋＤ、２９ ｋＤ、３３ ｋＤ、３５ｋＤ的多肽含量上升。蛋白质代谢的这种变化可能是植物的一种适应性反应     

激素对枸杞体细胞胚发生及可溶性蛋白质含量和组分的影响

以宁夏枸杞无菌苗叶片为材料,对体细胞胚发生过程中激素作用、可溶性蛋白质变化及体细胞胚发生频率进行了研究。发现不同激素处理、可溶性蛋白质含量、组分和体细胞胚发生频率均有一定的差异,三者之间存在着相关性。结果如下：（１）ＭＳ＿１、ＭＳ＿２和ＭＳ＿３三种培养基上继代的愈伤组织难以诱导形成体细胞胚,为非胚性愈伤组织;而从ＭＳ＿１、ＭＳ＿２和ＭＳ＿３分别转到ＭＳ＿０和ＭＳ＿４培养基上的愈伤组织体细胞胚发生频率高,属于胚性愈伤组织。其中以ＭＳ＿３转至ＭＳ＿０后的体细胞胚发生频率最高。（２）可溶性蛋白质ＳＤＳ－ＰＡＧＥ分析表明：非胚性愈伤组织有特异性蛋白质６７ｋＤ,胚性愈伤组织有特异性蛋白质３５ｋＤ和４４ｋＤ;蛋白质３３ｋＤ和６７ｋＤ受２,４－Ｄ调控,蛋白质４９ｋＤ、５７ｋＤ受６ＢＡ调控,而蛋白质３７ｋＤ、５１ｋＤ受２,４－Ｄ和６ＢＡ协同调控。     

莲子在光下萌发过程中叶绿体色素蛋白质复合体及光化活性的变化

实验研究了莲种子（ＮｅｌｕｍｂｏｎｕｃｉｆｅｒａＧａｅｒｔｎ．）在光下萌发过程中叶绿体色素蛋白复合体、多肽组成以及ＰＳⅡ光化活性的变化。结果表明,萌发到第６天的莲胚芽叶绿体,在ＳＤＳＰＡＧＥ凝胶柱中只分出两条色素带,分别为捕光叶绿素蛋白复合体（ＬＨＣⅡ）和自由色素（ＦＰ）。萌发到第８天的莲叶绿体可分出ＣＰⅠ,ＬＨＣⅡ１、ＬＨＣⅡ和ＦＰ。仍未检测到ＰＳⅡ反应中心复合物。多肽分析表明：未萌发和萌发到第２天的叶绿体中３０ｋＤ多肽的含量显著,尽管２７ｋＤ多肽在未见光时已开始合成,但其含量很少。随着照光时间增长,３０ｋＤ多肽含量逐渐减少,２７ｋＤ多肽的含量逐渐增多,到第１２天时超过３０ｋＤ多肽的含量。电子传递速率和室温荧光诱导动力学的测定表明：ＰＳⅡ光化活性在莲子萌发到第７天时才开始出现,并随萌发时间增加而升高,与此同时Ｃｈｌａ／ｂ比值由低到高达到正常值。莲子在光下萌发过程中其叶绿体膜成分及膜功能的变化与它的超微结构的变化相符合。结果再次证明,莲胚芽叶绿体在光下的发育具有与其它高等植物所不同的独特途径,这为莲在被子植物系统发育中占有独特地位的看法提供了依据。     

水分胁迫下玉米叶肉细胞超微结构的变化及其与膜脂过氧化伤害的关系

研究ＰＥＧ模拟水分胁迫条件对玉米叶肉细胞超微结构的影响,探讨了超微结构的变化与保护酶活性及膜酯过氧化伤害之间的关系。试验表明,在水分胁迫初期,叶肉细胞超微结构变化较小,此时,叶片的ＳＯＤ及过氧化氢酶活性明显升高,质膜相对透性和丙二醛含量增加缓慢,随着胁迫时间的延长,叶肉细胞超微结构破坏加重,且不同细胞对水分胁迫的敏感性相差很大,叶片ＣＡＴ性下降,质膜透性和ＭＤＡ含量急增;复水后,叶片超微结构,ＳＯ     

盐胁迫对大麦幼苗质膜,液泡膜上共价和非共价结合多胺含量的影响

用不同浓度ＮａＣｌ处理７ｄ龄大麦（Ｈｏｒｄｅｕｍ ｖｕｌｇａｒｅＬ．）幼苗３ｄ。以非共价键和共价键形式分别与质膜和液泡膜微囊及膜蛋白结合的多胺含量受低 度盐的促进而被高浓度盐所抑制。以非共价键形式与膜微囊结合的各种多胺中亚精胺（Ｓｐｄ）含量最高,占膜上多胺总量的４０％－７０％,与膜蛋白共价结合的各种多胺中腐胺（Ｐｕｔ）含量占主导地位,占膜蛋白上多胺总量的３５％－６０％。在根系液泡膜上发现一种含量丰     

水分胁迫对小麦根细胞质膜氧化还原系统的影响

水分胁迫使小麦根质膜ＮＡＤＨ和ＮＡＤＰＨ的氧化速率及Ｆｅ（ＣＮ）６＾３－和ＥＤＴＡ－Ｆｅ＾３＋的还原速率明显降低。对照与胁迫处理的质膜氧化还原系统活性均不受鱼藤酮、抗霉素Ａ和ＤＣＮ等呼吸链抑制剂的影响。在不加Ｆｅ（ＣＮ）６＾－３作为电子受体时,水杨基羟肟酸（ＳＨＡＭ）可明显刺激质膜ＮＡＤＨ的氧化和Ｏ２吸收速率。水分胁迫促使ＳＨＡＭ刺激的ＮＡＤＨ氧化明显降低,但却使Ｏ２吸收略有上升。     

磷酸饥饿时番茄幼苗酸性磷酸酶活性的变化与Pi吸收的关系

磷酸饥饿时,番茄幼苗根部及地上部酸性磷酸酶活性均显著增强,根部细胞表面酸性磷酸酶及根部外泌的酸性磷酸酶活性亦明显提高。动力学分析表明,磷酸饥饿提高了番茄幼苗根部的酸性磷酸酶对其底物的亲和力。另外,磷酸饥饿对番茄幼苗根部酸性磷酸酶活性的最适ｐＨ值没有影响。钼酸对番茄幼苗根部酸性磷酸酶活性有强烈的抑制作用,对番茄幼苗Ｐｉ吸收速率也有十分明显的抑制效果。以上结果表明,磷酸饥饿时,番茄幼苗Ｐｉ吸收的适应性变化可能与根部酸性磷酸酶特别是根部细胞表面酸性磷酸酶及其外泌酸性磷酸酶的参与密切关联。     

Relationship between the Changes of Acid Phosphatase Activities and Pi -uptake of Tomato Seedlings during Phosphate Starvation

The acid phosphatase activities from roots and both stems and leaves of tomato seedlings all in-creased markedly under phosphate starvation. Phosphate starvation also increased the activities of acid phos-phatase from cell surface of, and released by roots of tomato seedlings. The kinetic analysis of acid phos-phatase of roots of tomato seedlings revealed that phosphate starvation increased the affinity of the enzyme to its substrate. The results also revealed that phosphate starvation had no effect on the optimum pH (pH 4.93) of the acid phosphatase of roots of tomato seedlings. It was also found that molybdate strongly inhibited not only the activities of acid phosphatase but also Pi- uptake rates of tomato seedlings.     

磷饥饿时番茄幼苗根系质膜H+-ATP酶活性

磷饥饿提高了番茄幼苗质膜H+-ATP酶活性并促进了番茄幼苗根部的H+分泌。动力学分析表明,磷饥饿使番茄幼苗根部质膜H+-ATP酶的Km值明显降低,亦即提高了该酶对其底物的亲和力,但对该酶的Vmax影响不大。另外,磷饥饿并不改变ATP酶的最适pH值(最适pH值为6.5)。钒酸盐显著抑制番茄幼苗根部质膜ATP酶的活性以及H+分泌,也显著抑制番茄幼苗的Pi吸收。与对照相比,上述抑制作用在饥饿处理的植物中表现得更强。以上结果表明,磷饥饿时高亲和性Pi传递系统的诱导很可能包含质膜H+-ATP酶的参与。     

Quantitative changes in maize cytoplasmic proteins induced by aluminium

Three-day-old maize seedlings were subjected to 100 μM AlCl3 for 24 h. Cytoplasmic proteins were isolated from root tips, root base and from coleoptiles. After fractionation of cytoplasmic proteins on anion chromatography column Bio-Scale Q2 sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis was used to monitor Al-induced changes in polypeptide composition of particular fractions. Four (root) and 7 (coleoptile) fractions were eluted from the column with linear 0 - 1.0 M NaCl gradient. In fraction 1 of cytoplasmic proteins from root tips Al induced accumulation of polypeptide with molecular mass of 16 kD and simultaneous reduction of two polypeptides (67.5 and 60 kD). In fraction 1 isolated from mature zone of maize roots Al-induced accumulation of 22 kD polypeptide and reduction of 67.5, 60, and 14 kD polypeptides. Most pronounced changes were revealed in coleoptile. In three protein fractions increased accumulation of polypeptides with molecular mass of 14, 17.5, 20, 24.5, 28, 30, and 37.5 kD were observed. In the remaining three root or four coleoptile fractions of cytoplasmic proteins, no differences were found between Al-treated and control maize seedlings. This revised version was published online in July 2006 with corrections to the Cover Date.     

Electrophoretic characterization of a detergent-treated plasma membrane fraction from corn roots

Experiments were conducted to determine conditions essential for electrophoretic characterization of a detergent-extracted plasma membrane fraction from corn (Zea mays L.) roots. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) initially gave poor resolution of polypeptides in the plasma membrane fraction and, upon detergent treatment for purification of the proton-pumping adenosine triphosphatase (ATPase), showed no enrichment for a 100 kilodalton catalytic subunit characteristic of the ATPase. In contrast to SDS-PAGE, phenol urea acetic acid (PAU)-PAGE clearly resolved two polypeptides in the 100 kilodalton region that were enriched during detergent treatment and indicated at least one polypeptide forms a phosphorylated intermediate characteristic of the ATPase. Problems with SDS-PAGE were found to be caused, in part, by a combination of endogenous proteases and heat-induced aggregation of high molecular weight proteins. The usually standard procedure of boiling the sample prior to SDS-PAGE caused the aggregation of the 100 kilodalton polypeptides. By controlling for proteases using chymostatin and/or phenylmethane sulfonyl floride, and not boiling the sample prior to electrophoresis, two polypeptides were clearly resolved by SDS-PAGE in the 100 kilodalton region of Triton X-114-extracted membranes from corn, oat, barley, and tomato.     

Accumulation of Dehydrins and ABA-Inducible Proteins in Wheat Seedlings during Low-Temperature Acclimation

Using one-dimensional SDS-PAGE and immunochemical methods, we detected the presence and estimated the content of dehydrins and ABA-inducible (RAB) polypeptides in etiolated seedlings of four spring and three winter wheat (Triticum aestivum L.) cultivars differing in frost hardiness. We hardened three-day-old seedlings at 4°C for nine days or grew them at 22°C for a day (control seedlings). We established that heat-stable cold-regulated (COR) polypeptides with mol wts of 209, 196, 169, 66, 50, and 41 kD, which are characteristic of hardened wheat seedlings, were homologous to polypeptides from a dehydrin family and polypeptides with mol wts of 209, 196, 66, 50, and 41 kD were immunologically related to RAB-proteins. We supposed that these COR polypeptides were involved in the prevention of local protein dehydration and denaturation during hypothermia. Analysis of the relative content of COR proteins revealed a close correlation between the cultivar frost hardiness and the concentration of these proteins. It seems evident that different accumulation of dehydrins and RAB polypeptides in different cultivars of a single species is one of the causes for different plant frost hardiness.     

Effect of chilling temperatures on osmotic water permeability and aquaporin activity in the plasma membranes from pea roots

The osmotic water permeability of plasma membrane vesicles was examined after isolation from the roots of 7-day-old etiolated pea ( Pisum sativum, cv. Orlovchanin) seedlings grown at optimal temperature and those exposed to 1-day chilling at 8°C in the end of the growth period. The homogenization medium for obtaining plasma membranes was supplemented with either SH-reagents or protein phosphatase inhibitors. The plasmalemma vesicles were purified from the microsome fraction by means of two-phase polymer system. The osmotic water permeability of membrane vesicles was evaluated from the rate of their osmotically induced shrinkage. The lowering of growth temperature was accompanied by the increase in osmotic water permeability of plasmalemma. These changes occurred without the corresponding increase in aquaporin content or permeability of membrane lipid matrix. The membranes from cooled seedlings were markedly depleted in the content of SH-groups. Furthermore, the treatment of membrane samples with a thiol-reducing agent, tributylphosphine did not raise the SH-group content in membranes from chilled plants, unlike such changes in membranes from warm-grown plants. When the homogenization medium contained dithiothreitol and phenylarsine oxide (an inhibitor of tyrosine protein phosphatases), the osmotic permeability of plasmalemma in preparations from warm-grown seedlings also increased. Based on these results, it is supposed that aquaporin-mediated water permeability of membranes is regulated through different pathways under optimal and adverse conditions for plant growth. Direct action of endogenous SH redox regulators on aquaporin activity is likely under optimal growth conditions, while protein phosphatase might mediate changes in aquaporin activity under unfavorable growth conditions.     

Protein Kinases in Zucchini (Characterization of Calcium-Requiring Plasma Membrane Kinases)

Using an in situ phosphorylation assay with zucchini (Cucurbita pepo L. cv Dark Green) seedling tissue, we have identified numerous polypeptides that are capable of acting as protein kinases. Total protein preparations from different organs contain different kinase profiles, but all are within the range of 55 to 70 kD. At least four kinases are associated with highly purified plasma membranes from etiolated zucchini hypocotyls. The major phosphorylated polypeptides from plasma membranes range in apparent molecular mass from 58 to 68 kD. The plasma membrane kinases are activated by micromolar concentrations of calcium and phosphorylate serine, and, to a lesser extent, threonine residues. These characteristics are similar to those of a soluble calcium-dependent protein kinase that has been purified to homogeneity from soybean suspension cultures. Three of the zucchini plasma membrane kinases share antigenic epitopes with the soluble soybean kinase. The presence of kinase activity at different apparent molecular masses may be indicative of separate kinases with similar characteristics. The zucchini hypocotyl protein kinases are not removed from plasma membrane vesicles by 0.5 M NaCl/5 mM ethylenediaminetetraacetate or by detergent concentrations below the critical micelle concentration of two types of detergent. This indicates that the plasma membrane protein kinases are tightly associated with the membrane in zucchini seedlings.