萝卜颗粒引发大豆种子抗吸胀冷害能力的效应

以鲜萝卜颗粒和聚乙二醇(PEG)引发大豆种子抗吸胀冷害能力的研究结果表明,二者都能引发和控制种子在低温吸胀过程中的吸水,提高种子活力和萌发率,减少细胞中电解质和氨基酸外渗。     

花生种子萌发吸胀阶段冷害抗性的鉴定及耐冷种质的筛选

为更好地评价花生吸胀冷害抗性,本研究首先分析了常温条件下(25℃)和冷胁迫条件下(2℃)花生种子萌发过程中的吸水状况,结果表明不同基因型的花生吸胀阶段的持续时间基本一致,维持在浸种后0～12 h;冷胁迫能够使吸胀阶段初期的吸水速度显著下降,但对吸胀阶段的持续时间没有显著影响。对种子萌发3个生理阶段(吸胀、萌动、萌发阶段)分别进行冷害胁迫处理,结果表明吸胀阶段遭受的冷害胁迫对花生种子萌发影响较其他阶段严重。进一步的试验表明,吸胀阶段种子2℃冷胁迫12 h,能够更好地区分不同花生种质之间吸胀冷害抗性的差异,从而建立了花生种子吸胀冷害抗性的鉴定体系。利用该鉴定体系对64份花生种质的吸胀冷害抗性进行鉴定,以相对发芽率85%为阈值,筛选出了7份耐冷种质。本研究为花生耐冷育种及吸胀冷害抗性机制解析提供了技术鉴定体系和材料基础。     

渗透调节对冷敏感大豆[Glycine max(L.)Merr.]种子膜类脂组成和脂肪酸含量的影响

选用冷敏感的大豆[Gzycine max（L.）Merr．]品种“中黄22”为试材,运用薄层层析法和气相色谱法研究了渗控增强大豆种子活力过程中膜类脂的组成及变化规律,揭示生物膜类脂在抗吸胀冷害中的作用。结果表明,用33％（W／V）的PEG6000处理种子,随着渗控时间的延长,种子的发芽指数、活力指数和根干重都逐渐增加,在渗控72h时,大豆种子中不饱和磷脂如PC、PE的比例增加,各极性脂的脂肪酸组成也发生了变化,PC、PE和PI中饱和脂肪酸16：0的含量显著下降,而不饱和脂肪酸18：2明显上升,使它们的不饱和指数大大提高,并与渗控时间呈正相关。由以上结果可证明,经PEG渗控处理后,提高了冷敏感大豆膜的流动性,为防止吸胀冷害,保持膜系统的完整性和生物膜正常的代谢提供了前提条件。     

PVA和PEG预处理对大豆种子在低温吸胀过程中胚根线粒体发育和超微结构的影响

电镜观察发现,大豆种子在刚开始萌发时胚根细胞中未能见到线粒体,线粒体是在种子萌发过程中逐渐出现的,由原质体再分化发育而成。对照胚根细胞内原质体在低温吸张过程中明显膨胀,在回温后胚根细胞中原质体仍不能发育成线粒体,甚至网状膜结构破坏,呈空泡化;经聚乙烯醇(PVA)和聚乙二醇(PEG 6000)预处理的大豆种子在同样条件下线粒体能继续发育,在回温后预处理胚根细胞中线粒体发育良好,具有明显的双层膜和管状嵴的结构。这些结果表明,在低温吸胀过程中原质体能够继续再分化发育成线粒体是提高大豆种子活力和抗冷力的重要原因。     

聚乙二醇(PEG)引发预防大豆种子吸胀冷害的效果

PEG引发法是一种预防敏感大豆种子吸胀冷害的有效方法。我们首次发现引发种子能抗御长达2个月的2～3℃低温而不降低种子活力,由之发育的幼苗其离体子叶在普通滤纸发芽床上形成愈伤组织并有再分化现象。引发过程中种子呼吸强度逐渐提高,推测PEG引发提高抗性的作用与种子代谢活动的预先启动有关。     

烟草种子成熟过程中膜脂的变化规律研究

该研究采用脂类组学方法,系统地研究了烟草种子成熟过程中膜脂含量及组成比例的变化规律。结果表明:(1)构成叶绿体和类囊体膜的重要脂类质体膜脂的含量及其在总膜脂中的组成比例,在种子成熟的整个过程中保持下降趋势;而构成细胞膜的重要脂类质外体膜脂含量在种子成熟前期则下降显著,在授粉21 d后基本保持不变。(2)总膜脂含量的变化规律与质体膜脂类似,但在授粉后第29天后含量却达到稳定状态。(3)因油脂在种子成熟过程中不断积累,且化学结构与膜脂相似,质体膜脂含量的降低可能与种子成熟过程中种子对油脂累积的持续需求以及对叶绿体及类囊体的需求降低有关。(4)质外体膜脂含量在授粉21 d后基本保持不变的原因,可能是由于脂质外体膜脂是细胞膜组成的主要膜脂,细胞膜在种子成熟以及成熟种子萌发过程中均发挥重要作用,因此质外体膜脂只在种子成熟的前期有部分转化为油脂。     

一氧化氮对番茄种子抗吸胀冷害的影响

以番茄毛粉802种子为材料,通过对比实验,测定分析各处理种子的萌发率及第4天的平均根长、萌发指数、活力指数,以及相对电导率(REC)、丙二醛(MDA)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)含量的变化,以探讨NO对番茄种子吸胀冷害的抵抗作用及其机理.结果显示:(1)外源NO可显著提高番茄种子经12 h吸胀冷害处理后的萌发率、平均根长、萌发指数和活力指数,并显著降低吸胀冷害下REC和MDA含量,同时显著提高SOD和CAT的含量.(2)NO所提高的吸胀冷害处理后种子的SOD和CAT活性不能被RNA合成抑制剂放线菌素D和蛋白质合成抑制剂环己酰亚胺抑制.结果表明,NO可提高番茄种子抵抗吸胀冷害的能力,而且与NO激活了抗氧化系统有关,但NO不是通过促进抗氧化酶的合成来提高其活性.     

PEG预处理对早稻种子活力和抗寒能力的影响(简报)

应用25％PEG在25℃下预处理48h,可以明显地提高早稻种子活力和抗寒能力,减少5℃低温吸胀过程中的离子渗漏,提高萌发种子的淀粉酶活力及种子可溶性糖和可溶性蛋白质含量。在早稻育秧中有一定的应用价值。     

拟南芥叶片衰老过程中细胞溶血磷脂和膜脂不饱和度的变化

细胞膜的流动性和渗透性的改变是植物衰老过程中一个内在的、具有破坏性的变化。膜脂组成中,溶血磷脂的出现是膜伤害的一个重要标志;膜脂双键数目的变化是影响膜流动性的主要因素。应用脂类组学的方法,检测了拟南芥野生型及其磷脂酶Dδ (PLDδ)缺失型突变体在离体诱导的、脱落酸（abscisic acid, ABA）和乙烯（ethylene）促进的衰老过程中,溶血磷脂（lysophospholipids, lysoPLs）的分子变化,并通过计算膜脂双键指数（double bond index, DBI）表征了膜流动性的变化。结果表明,在离体诱导的衰老过程和乙烯促进的衰老过程中,溶血磷脂的总含量和各溶血磷脂分子的变化不显著,而在ABA促进的衰老过程中溶血磷脂总含量和部分溶血磷脂分子均显著升高;在上述三种衰老处理下,总膜脂的DBI均下降,但是离体诱导和激素促进的的衰老过程中各类膜脂的DBI的变化却不同。同时我们还发现,抑制PLDδ基因表达降低了ABA促进的衰老过程中溶血磷脂的产生、减缓了ABA和乙烯促进的衰老过程中总的膜脂的DBI的降低。     

高压静电场（HVEF）对大豆种子吸胀冷害的影响

高压静电场预处理可不同程度地提高大豆种子抗低温吸胀冷害的能力,处理组种子在低温吸胀时无机离子,可溶性糖和可溶性蛋白的外渗量显著低于对照组,静电场预处理还提高了种子中过氧化氢酶（简ＣＡＴ）、过氧化物酶（简ＰＯＤ）、超氧物歧化酶（简ＳＯＤ）等清除自由基的酶活性,降低了丙二醛（简ＭＤＡ）的积累,促进了种子代谢水平,即提高了种子的淀粉酶、脱氢酶的活力和可溶性蛋白含量,最终提高了发芽率。本文以膜的相变和损伤     

How membranes organize during seed germination: three patterns of dynamic lipid remodelling define chilling resistance and affect plastid biogenesis

Imbibitional chilling injury during germination causes agricultural losses, but this can be overcome by osmopriming. It remains unknown how membranes reorganize during germination. Herein, we comparatively profiled changes of membrane lipids during imbibition under normal and chilling temperatures in chilling‐tolerant and ‐sensitive soybean seeds. We found three patterns of dynamic lipid remodelling during the three phases of germination. Pattern 1 involved a gradual increase in plastidic lipids during phases I and II, with an abrupt increase during phase III. This abrupt increase was associated with initiation of photosynthesis. Pattern 3 involved phosphatidic acid (PA) first decreasing, then increasing, and finally decreasing to a low level. Patterns 1 and 3 were interrupted in chilling‐sensitive seeds under low temperature, which lead a block in plastid biogenesis and accumulation of harmful PA, respectively. However, they were rescued and returned to their status under normal temperature after polyethylene glycol osmopriming. We specifically inhibited phospholipase D (PLD)‐mediated PA formation in chilling‐sensitive seeds of soybean, cucumber, and pea, and found their germination under low temperature was significantly improved. These results indicate that membranes undergo specific and functional reorganization of lipid composition during germination and demonstrate that PLD‐mediated PA causes imibibitional chilling injury.     

拟南芥角果衰老过程中膜脂的变化

角果发育对某些物种的生殖发育具有重要的作用。拟南芥种子附着在角果里,角果在早期发育时进行光合作用,角果成熟后开裂散落种子之前,其细胞会经历一个衰老的过程。一般植物细胞在衰老过程中要经历膜脂降解的过程,但是角果细胞衰老过程仍未知。通过比较角果衰老过程中拟南芥野生型(WS)及与膜脂代谢密切相关的磷脂酶Dδ缺失突变体(PLDδ KO)中膜脂分子的组成情况、膜脂含量、相对含量及双键指数值,结果发现,在拟南芥角果衰老过程中：(i)质体膜脂和质体外膜脂显著下降;(ii)不同膜脂降解速率不一样,质体膜脂的降解比质体外膜脂的降解快;(iii)总的双键指数DBI下降;(iv)磷脂酶Dδ缺失突变体(PLDδ KO)的角果膜脂组成的基本水平和变化样式与野生型(WS)非常相似。结果说明,角果在衰老过程中发生了膜脂的激烈降解。据此推测：(i) 膜脂水解产物可能转移到种子中用于储藏脂三酰甘油的合成;(ii) 质体膜脂相对含量下降和质体外膜脂相对含量上升导致了总的DBI下降;(iii) PLDδ参与了角果衰老中的膜脂代谢。     

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.     

Effect of Low Temperature lmbibition on Mito chondrium Respiration and Phosphorylation of PEG Primed Soybean Seed

Cotyledon mitochondrium respiration and oxidative phosphorylation activity of PEG primed and unprimed (control) soybean seeds which have been exposed to low temperature imbibition before germination are studied. The ADP stimulated respiration rates of control mitochondria are evidently higher than state Ⅲ respiration rates of mitochondria from primed seed when L-Mal, α-Kg and Succ are used as substrates respectively. The mitochondria from the unprimed do not possess respiratory control (RC.) On the contrary, mitochondria from the primed, even after seeds being exposed to 2–3 ℃ imbibition for 24 h, phosphorylate normally. The ADP/O and RC values are consistent with those of theoretical expectation. When NADH is used as substrate, unprimed seed mitochondria still possess oxidative phosphorylation activity, while ADP/O and RC values are obviously lower than those of mitochondria from the primed. The emerging sequence of the activity of the diverse phosphorylation sites during germination is also studied. When a different substrate is used, the emerging sequence of the primed is as follows: 1. NADH (12 h), 2.α-Kg (24 h), 3. L-Mal and Succ (48 h). This corresponds to occurrence sequence of ADP stimulated respiration in control mitochondria. The above results show that low temperature imbibition has an irreversible destructive effect on oxidative phosphorylation activity of control mitochondria, and PEG priming has a protective effect on structure and function of the mitochondria under low temperature imbibition stress. The mechanism of soybean imbibitional chilling injury and protective effect of PEG priming are discussed.     

Physiological Effects of Hydration-Dehydration Treatment on Soybean Seeds to Enhance Its Resistance to Imbibitional Chilling Injury

The soybean (Glycine max Merrill) cv. “Heihe No. 3” which is sensitive to imbibitional chilling injury was used as experimental material. From the point of cell membranes and by means of physiological tests, the relation of hydration-dehydration to membrane repairation during rehydration in seeds was discussed, and a new point of view on the acting mechanisms of hydration-dehydration was approached. The resistance of seed to imbibitional chilling injury was obviouslv enhanced, its apparent efficiency displayed the enhance of seed rigor, reduction of leakage, especially the K+/Na+ ratio; increasing the K+-Na+-ATPase activity,. The facts indicate that the function of cytoplasmic membrane was enhanced and maintained normally. In the meantime, the activities of SOD, catalase and peroxidase were all enhanced, and the production of volatile aldehydes were decreased. It shows that the function of cell antiautoxidation system was greatly improved. Based on a series of physiological and biochemical index mentioned above and the comparison of water-absorbed curves, a postulated model of membrane system induced by hydration-dehydration treatment is given. It is considered that the effects of the two processes, i,e. physical membrane repairation as well as the physiological and biochemical repairation, will be persisted in the redehydrated seeds.     

Effects of PVA and PEG Pretreatment on Development and Ultrastructure of Plumular Root Mitochondria in Soybean Seed During Low Temperature Imbibition Process

Electron microscope observation revealed that mitochondria in plumular root cells of soy bean seed were invisible at the beginning of seed germination. They were sequentially formed re-differentiated and developed from proplastids during the process of germination. The proplastids in the control plumular root markedly expanded at low temperature imbibition process and did not develop into mitochondria when normal temperature was restored. Even the structure of reticulata membrane was further destroyed and vacuolized. In contrast, mitochondria of soybean seed pretreated with polyvinyl alcohol (PVA) and polythylene glycol-6000 (PEG) continused to develop well under the same conditions, as characterized by a clear structure of bilayered membrane and tube cristae. The results suggest that continuous re-differentiation and development of proplastids into mitochondria at low temperature imbibition process play an important role in the increase of soybean seed vigour and its resistance to chilling injury.     

Ascorbate-glutathione cycle of mitochondria in osmoprimed soybean cotyledons in response to imbibitional chilling injury

Osmopriming treatment of chilling-sensitive soybean (Glycine max L. cv. Zhonghuang-22) seeds for 72 h at 25 °C with polyethylene glycol (PEG8000) solution at −1.5 MPa strongly improves chilling resistance. The aim of the present work was to investigate whether the beneficial effect of osmopriming is associated with restoration of the ascorbate-glutathione (ASC-GSH) cycle of mitochondria in soybean seeds. Compared with the control, both H₂O₂ and malondialdehyde (MDA) contents in mitochondria of osmoprimed seeds decreased after chilling treatment, and these changes were associated with increased activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and l-galactono-γ-lactone dehydrogenase (GLDH). However, the activity of dehydroascorbate reductase (DHAR) showed no obvious change during osmopriming treatment. Increased ASC and GSH contents accompanied prolonged osmopriming, and the reduced/oxidized ratios of ASC and GSH increased differently during osmopriming. These results indicate that osmopriming treatment enhances activity of the ASC-GSH cycle of mitochondria, which raises the chilling tolerance in soybean seeds and protects against H₂O₂ that is generated in mitochondria during imbibition at low temperature.