花生种子萌发过程中胚轴多胺氧化酶的活性变化

研究花生（Ａｒａｃｈｉｓｈｙｐｏｇａｅａ）种子萌发过程中胚轴多胺氧化酶（ＰＡＯ）的活性变化及其与种子萌发的关系表明：胚轴中的ＰＡＯ活性是在种子萌发过程中逐渐形成的,而黑暗条件更有利于该酶的活性形成;放线菌素Ｄ（１０ｍｇ／Ｌ）、环己酰亚胺（１０ｍｇ／Ｌ）处理对种子萌发的抑制率分别为２６．３％和８７．３％,对胚轴ＰＡＯ活性的抑制率分别为４１．１％和９４．０％,显示胚轴中的ＰＡＯ很可能参与花生种子的萌发过程,且其ｍＲＮＡ在种子发育过程中已合成并贮存于种子中,萌发时ＰＡＯ活性的出现主要是由于这些ｍＲＮＡ转译合成了ＰＡＯ。     

大豆初生幼苗多胺氧化酶活性的细胞化学定位

对大豆(Glycinemax(L.)Merril“l)垦农4号”萌发种子和初生幼苗中的多胺氧化酶(polyamineoxidase,PAO,EC1.4.3.6)的活性和分布进行了研究。结果表明,PAO活性仅在种子萌发起始后(吸胀后24h)才检测到,然后随着种子萌发进程,PAO活性快速升高。但是,在萌发种子(吸胀后72h)和初生幼苗(吸胀后120h)中,PAO活性在各器官中的分布有明显差异。在萌发种子中,PAO活性在胚根最高(5.17±0.91Ug-1FW),胚轴次之,胚芽再次之,子叶活性最低(0.12±0.03Ug-1FW);在初生幼苗中,PAO活性在下胚轴中最高(5.47±0.66Ug-1FW),幼根次之,顶芽再次之,子叶最低(0.10±0.03Ug-1FW)。这种差异对种子萌发和幼苗形态建成有积极意义。运用细胞化学定位在透射电镜下观察初生幼苗PAO在各部位的分布,发现PAO主要定位在顶芽细胞的液泡膜上、子叶细胞的细胞壁及其外侧表面、下胚轴细胞的细胞壁及其表面,且PAO与细胞壁表面结合较紧;根细胞的细胞壁、细胞间隙、细胞膜、液膜上均有分布,但以液泡膜分布居多。本研究结果进一步证实了PAO在细胞壁和细胞间隙有着较广泛的分布。首次报道PAO在细胞膜和液泡膜上有分布。     

发育和早萌中花生胚胚轴与子叶内BAPAase活性的差异

花生胚发育过程中,子叶和胚轴中都出现BAPAase活性。花生种子萌发时,子叶和胚轴中的BAPAase活性迅速上升,子叶中无新的BAPAase合成,胚轴中能重新合成BAPAase。ABA抑制了子叶和胚轴中BAPAase的活性,抑制胚轴中BAPAase活性所需的外源ABA浓度更高,Act-D和CHM不能逆转ABA对BAPAase活性的抑制作用。     

光活化α-三噻吩诱发稗草愈伤组织保护酶系的反应

用α-三噻吩(α-T)处理稗草(Echinochloa crusgalli)愈伤组织,经近紫外光照射后,形成细胞内的氧化胁迫环境。发现经0．1、1和10mg／L浓度α-T处理,所测谷胱甘肽-S-转移酶(GST)、谷胱甘肽过氧化物酶(GSH-Px)和过氧化物酶(POD)的光照诱导活性程度明显高于非光照的活性,其中1mg／L浓度处理,光照所提高活性分别为26.78％、217．66％和124．72％。但随着处理浓度的提高,超氧化物歧化酶(SOD)的活性呈下降趋势,以1mg／L和10mg／L的浓度处理,所测SOD抑制率分别为19．95％和55．44％。     

去胚轴对花生子叶肽链内切酶和贮藏蛋白质降解的影响

从离体子叶与连体子叶在水中培养一段时间后的比较,看到它们之间在肽链内切酶活性和盐溶蛋白及花生球蛋白降解上的差异并不大,这表明去除胚轴对子叶肽链内切酶活性和贮藏蛋白降解的影响很轻微。亚胺环己酮(蛋白质合成抑制剂)不能完全抑制离体子叶肽链内切酶活性的提高,子叶的大部分大分子贮藏蛋白同样被降解。这表明,在花生种子萌发过程中降解大部分贮藏蛋白的子叶肽链内切酶并非全部是在种子萌发时新合成的,子叶贮藏蛋白降解和肽链内切酶活性基本不受胚轴调控,子叶与胚轴之间在调控关系上可能是一种新的调节类型。     

花生种子萌发早期胚轴DNA合成与种子活力的关系

随着花生种子萌发率和活力指数的下降,胚轴DNA开始合成的时间推迟,其合成水平也降低。但DNA合成都是先于吸胀的12h(高活力胚)或18h(低活力胚)出现一个峰,然后再持续上升。腐胺预处理明显地促进老化胚轴萌发早期(12～2dh)的DNA合成。钙离子预处理则有一定抑制作用,但两种预处理均能提高种子的活力指数,并能促进吸胀30h以后的DNA合成。     

发育和早萌中花生胚胚轴一子叶内BAPAase活性的差异

花生胚发育过程中,子叶和胚轴中都出现ＢＡＰＡａｓｅ活性,花生种子明发时,子叶和胚轴中的ＢＡＰＡａｓｅ活性迅速上升,子叶中无新的ＢＡＰＡａｓｅ合成,胚轴中能重新合成ＢＡＰＡａｓｅ。ＡＢＡ抑制子叶和胚轴中ＢＡＰＡａｓｅ的活性,抑制胚轴中ＢＡＰＡａｓｅ活性所需的外源ＡＢＡ浓度更高,Ａｃｔ＝Ｄ和ＣＨＭ不能逆ＡＢＡ对ＢＡＰＡａｓｅ活性的抑制作用。     

不同成熟度花生胚萌发时子叶中贮藏蛋白质的降解

花生(Arachis hypogaea L.)汕油71果针入土20d(20 DAP)的种子剥去种皮后,10％的胚可以萌发,至40 DAP发芽率达98％。不同发育时期的花生胚萌发 10d后子叶盐溶蛋白质和花生球蛋白降解表明,20和32 DAP胚萌发后,子叶中这些蛋白质只有部分降解。随着胚成熟度增加,子叶中降解这些蛋白质的能力不断提高。20～40 DAP胚萌发4d时,子叶的BAPAase和GHE活性较低。50～80DAP胚萌发 4d,子叶中上述两种酶均显示较高的活性。     

脱落酸对发育中花生胚萌发和贮藏蛋白质合成的影响

发育中的花生胚在无激素固体培养基上高体培养时提前萌发,其发芽力随胚的成熟增加而提高。果针入土后40d胚的发芽率达100％。禹体培养过程中,外源ABA能够阻止花生胚提前萌发和促进胚的发育。胚成熟前期,较低浓度的ABA(10~(-5)mol/L)便抑制胚的萌发;而在成熟中期以后,则要求较高浓度的ABA(10~(-4)mol/L)才能抑制胚的萌发。ABA对成熟前期胚的贮藏蛋白质合成无影响,而对成熟中期至后期胚的贮藏蛋白质合成起促进作用。ABA维持花生胚贮藏蛋白质合成和积累的作用表现在转录水平上。     

一株放线菌代谢产物除草活性的初步研究

从土壤中分离筛选出1株编号为Z40的链霉菌菌株,其代谢产物具有明显的除草活性。室内生物测定结果表明,Z40菌株发酵液干物质的除草活性主要表现为抑制生长作用。对反枝苋的主根和主茎生长抑制中浓度(EC50)分别为276.76 mg/L和1 609.37 mg/L,对狗尾草的主根和主茎生长抑制中浓度分别为501.81mg/L和629.01 mg/L。浓度为10 000 mg/L对油菜的萌发抑制率和主茎生长抑制率均为100.00%,而对小麦的生长发育没有明显影响。田间试验结果表明,Z40菌株代谢产物对稗、狗尾草、铁苋菜的防治效果均在80%以上,抑制率分别为80.30%,81.59%和88.71%。     

Osmopriming-Regulated Changes of Plasma Membrane Composition and Function were Inhibited by Phenylarsine Oxide in Soybean Seeds

The aim of the present work was to investigate the effects of osmoconditioning on chilling injury in chilling-sensitive soybean ( Glycine max (L.) Merr. Zhonghuang No. 22) seeds during imbibition. Low temperatures reduced the germination rate and no seed germinated at 1 °C. Osmoconditioning of seeds at 20 °C with a polyethylene glycol-8000 (PEG8000) solution at 1.5 MPa for 72 h followed by drying back to their initial moisture content (MC) reduced their chilling sensitivity. The phenylarsine oxide (PAO), an inhibitor of protein tyrosinephosphatases, was used to investigate the possible involvement of phosphorylation-dephosphorylation of Tyr residues in the plasma membrane composition and function when seeds were osmoconditioned. The results showed the germination of osmoconditioned seeds decreased significantly when PAO was added in PEG solution after chilling treatment. PAO inhibited changes in composition of plasma membrane phospholipids and fatty acid induced by osmocondition, indicated that tyrosine protein phosphorylation is involved in the regulatory mechanisms of osmocondition-responsive chilling in soybean seeds. Western blot result further indicated that osmocondition treatment improved the activity of plasma membrane H⁺-ATPase after chilling treatment, but this effect was abolished by PAO. The possible regulation mechanism by Tyr protein phosphorylation is discussed.     

Studies on the Peanut Seed Vigor Stored in Different Gases

Peanut seeds (water content 6.21% ) were stored at 38–40 ℃ for artificial ageing storage with different gases. N2 or CO2 delayed peanut seed ageing in comparison with air. The rate of seed germination was not affected by storing for 26 weeks under N2 or CO2, but the vitality of seed lost when stored under air. Determination of peanut seed vigor (root length+ axial length)×percentage of germination may precisely show the change in seed quality during ageing. The results indicate the positive correlation between the respiration and subsequent growth in peanut seeds, and the significantly negative correlation between vigor and electrical conductivity or sugar content in the seed extract.     

A Bacterial Indole-3-acetyl-L-aspartic Acid Hydrolase Inhibits Mung Bean (Vigna radiata L.) Seed Germination Through Arginine-rich Intracellular Delivery

Indole-3-acetyl-L-aspartic acid (IAA-Asp) is a natural product in many plant species and plays many important roles in auxin metabolism and plant physiology. IAA-Asp hydrolysis activity is, therefore, believed to affect plant physiology through changes in IAA metabolism in plants. We applied a newly discovered technique, arginine-rich intracellular delivery (AID), to deliver a bacterial IAA-Asp hydrolase into cells of mung bean (Vigna radiata) seeds and measured its effects on mung bean seed germination. IAA-Asp hydrolase inhibited seed germination about 12 h after the enzyme was delivered into cells of mung bean seeds both covalently and noncovalently. Mung bean seed germination was delayed by 36 h when the enzyme protein was noncovalently attached to the AID peptide and longer than 60 h when the enzyme protein was covalently attached to the AID peptide. Root elongation of mung bean plants was inhibited as much as 90% or 80%, respectively, when the IAA-Asp hydrolase was delivered with the AID peptide by covalent or noncovalent association. Further thin-layer chromatography analysis of plant extracts indicated that the levels of IAA increased about 12 h after treatment and reached their peak at 24 h. This result suggests that IAA-Asp hydrolase may increase IAA levels and inhibit seed germination of mung bean plants and that the AID peptide is a new, rapid, and efficient experimental tool to study the in vivo activity of enzymes of interest in plant cells.     

Evidence of a role for tyrosine dephosphorylation in the control of postgermination arrest of development by abscisic acid in Arabidopsis thaliana L

In the present paper evidence is presented indicating that tyrosine dephosphorylation is a key regulatory mechanism in postgermination arrest of Arabidopsis thaliana L. seed development mediated by abscisic acid (ABA). By using phenylarsine oxide (PAO), an inhibitor of tyrosine phosphatases, the sensitivity to the inhibitory effect of ABA on seed germination is enhanced. Consistent with this finding, we demonstrate that the ABA-responsive gene, RAB18, is hyperinduced in seeds imbibed in ABA plus PAO, compared with seeds imbibed only with ABA.     

Ethylene as a Component of the Emanations From Germinating Peanut Seeds and Its Effect on Dormant Virginia-type Seeds

The embryonic axes of Spanish-type peanut seeds that do not exhibit dormancy to any extent were found to produce ethylene during germination. Virginia-type peanut seeds of the extremely dormant variety NC-13 produced low levels of ethylene when imbibed but not germinating. Treatments that released dormancy of NC-13 peanut seeds resulted in increased ethylene production by the embryonic axis. The estimated internal concentration of ethylene in Virginia-type peanut seeds was 0.4 ppm at 24 hr of germination. Fumigation with an external concentration of 3.0 to 3.5 ppm for 6 hr was sufficient to break dormancy of Virginia-type peanut seeds. These results suggest that ethylene is associated with the germination processes of non-dormant seeds and participates in the breaking of seed dormancy of dormant peanut varieties.     

三种源地益智种子贮藏与萌发特性

利用0.4% 2,3,5-氯化三苯基四氮唑染色测定种子活力,研究广东、广西、海南三地栽培益智(Alpinia oxyphylla)种子的贮藏特性与萌发特性。结果表明,低温保湿条件有利于益智种子贮存,短期贮存可达150 d。益智种子发芽时间较短,发芽相对集中的最适条件是温度30 ℃、500 mg·L-1 GA3浸泡24 h;三个种源地的种子发芽率为88%～90%,不同种源地之间的种子萌发率没有显著差异。     

Characterization of isoperoxidase activity in Arachis hypogaea cultivars

Isoperoxidases (both anodic and cathodic) from individual seeds of several peanut cultivars (Arachis hypogaea), after ammonium sulfate precipitation of the major storage protein, arachin, from the extracts were characterized by polyacrylamide gel electrophoresis with respect to seed development, maturity and germination and the geographic areas where grown. All cultivars had a major cathodic isozyme near the origin of the gels. Much anodic intra- and inter-varietal isozyme polymorphism was observed in mature seeds collected from different geographic areas. These polymorphic isozymes were consistently present during the earlier stages of seed development (the activities decreased quantitatively and became variable during the later stages of maturation), and most were observed in each peanut upon germination, the latter showing quantitative increases in activity.     

Thiamine binding and metabolism in germinating seeds of selected cereals and legumes.

The basic characteristics of thiamine metabolism in germinating seeds of maize (Zea mays), oat (Avena sativa), faba bean (Vicia faba) and garden pea (Pisum sativum) are presented with a special emphasis of a possible thiamine storage function of seed thiamine-binding proteins (TBPs). Seeds were germinated for 6 d in the dark. Thiamine-binding activity in seeds decreased during germination by 50% in cereals and by 30% in legumes. The degradation of TBPs was also detected by polyacrylamide gel electrophoresis. The total thiamine content decreased rapidly to 20-40% of the initial value in cereal seeds during first 3 d of germination while in legume seeds thiamine content started changing from the fourth day and dropped by 50% at the sixth day. A composite pattern was found for the changes in thiamine pyrophosphate (TPP) contribution to total thiamine during seed germination. A peak of the coenzyme percentage was usually detected at the second day of germination. Another gain of TPP was often seen toward the sixth day of germination. The activity of thiamine pyrophosphokinase (EC 2.7.6.2) was high in resting legume seeds and did not significantly change during germination. In contrast, the low activity of this thiamine-activating enzyme in cereal seeds progressively increased during germination. Thiamine phosphate synthase (EC 2.5.1.3) was also detected in seeds and was shown to contribute significantly to the balance of thiamine compounds during seed germination.