水稻叶片中活性甲基循环、转移相关基因对干旱胁迫的应答

 以中旱3号、汕优63和矮仔占为材料,在10% PEG6000模拟干旱条件下,分别应用基因芯片和mRNA 差异显示技术检测了不同基因型水稻叶片中活性甲基循环、转移相关基因表达对干旱胁迫的应答。中旱3号和汕优63叶片中活性甲基循环过程受干旱胁迫诱导,而矮仔占叶片中这一过程受到干旱胁迫的抑制;干旱胁迫诱导中旱3号叶片中更多甲基转移酶基因的表达,尤其是对Rubisco蛋白甲基化修饰基因转录的诱导,有利于防止Rubisco蛋白的氧化降解;干旱胁迫抑制矮仔占叶片中DNA甲基转移酶、组蛋白甲基转移酶基因的转录。这一结果证明水稻中甲基循环、转移基因的表达在水稻耐旱中有一定的作用。
     

Effects of 1,2,4-Trichlorobenzene and Mercury Ion Stress on Ca^2＋ Fluxion and Protein Phosphorylation in Rice

The effects of 5 mg/L 1,2,4-trichlorobenzene (TCB) and 0.1 mmol/L mercury ion (Hg2 ) stresses on Ca2 fluxion and protein phosphorylation in rice seedlings were investigated by isotope exchange kinetics and in vitro phosphorylation assay. The Ca2 absorption in rice leaves and Ca2 transportation from roots to leaves were promoted significantly in response to Hg2 and TCB treatments for 4-48 h. The Ca2 absorption peaks presented in the leaves when the rice seedlings were exposed to Hg2 for 8-12 h or to TCB for 12-24 h. Several Ca2 absorption peaks presented in the roots during rice seedlings being exposed to Hg2 and TCB, and the first Ca2 absorption peak was at 8 h after being exposed to Hg2 and TCB. The result of isotope exchange kinetic analysis confirmed that short-term (8 h) Hg2 and TCB stresses caused Ca2 channels or pumps located on plasmalemma to open transiently. The phosphorylation assay showed that short-term TCB stress enhanced protein phosphorylation in rice roots (TCB treatment for 4-8 h) and leaves (TCB treatment for 4-24 h), and short-term (4-8 h) Hg2 stress also enhanced protein phosphorylation in rice leaves. The enhancement of protein phosphorylation in both roots and leaves corresponded with the first Ca2 absorption peak, which confirmed that the enhancement of protein phosphorylation caused by TCB or Hg2 stress might be partly triggered by the increases of cytosolic calcium. TCB treatment over 12 h inhibited protein phosphorylation in rice roots, which might be partly due to that TCB stress suppressed the protein kinase activity. Whereas, Hg2 treatment inhibited protein phosphorylation in rice roots, and Hg2 treatment over 12 h inhibited protein phosphorylation in rice leaves. This might be attributed to that not only the protein kinase activity, but also the expressions of phosphorylation proteins were restrained by Hg2 stress.     

镉胁迫对不同水稻品种种子萌发特性的影响

【目的】为了研究不同水稻品种的镉胁迫抗性差异,为早期预测镉污染,选育镉低积累水稻提供参考。【方法】采用50个不同遗传背景的水稻品种(系)种子,研究了0(蒸馏水,对照)、0.5、1.0、2.0 mmol/L等4种不同镉浓度胁迫对不同水稻品种种子发芽率、发芽指数、活力指数、胚芽长、胚根长、胚芽鲜质量与胚芽干质量等性状的影响。【结果】镉胁迫对水稻种子的萌发及幼苗生长具有抑制作用,且随着镉胁迫浓度的增高,抑制作用逐渐加强。0.5 mmol/L Cd~(2+)胁迫浓度对各水稻品种种子的发芽率和发芽指数影响不显著,而对其种子活力指数、胚根与胚芽生长则具有显著的抑制作用;不同镉胁迫浓度对水稻种子胚根的抑制作用明显大于胚芽;1.0mmol/L和2.0 mmol/L Cd~(2+)两种胁迫浓度处理下,不同水稻品种种子萌发期的各性状指标间差异达极显著水平,且达到极显著正相关。不同水稻品种对不同镉胁迫浓度的耐性存在明显差异。【结论】根据不同水稻品种萌发期不同镉胁迫浓度处理的各性状指标与对照的相对值进行聚类分析,可将50个供试水稻品种(系)分为镉胁迫敏感型、中间型和耐受型等3种类型。     

水分胁迫下外源多胺对水稻叶片光合速率与籽粒充实的影响

 以汕优63和PC311/早献党18为材料,研究了不同程度土壤干旱胁迫下水稻叶片中多胺含量变化以及外源多胺对籽粒充实和粒重的影响。在水分胁迫条件下,叶片中腐胺含量和腐胺与亚精胺以及精胺的比值提高。无论在土壤水分充足或水分胁迫条件下,抽穗期施用低浓度Spd和Spm后,叶片中的腐胺和MDA含量降低,而亚精胺和精胺、光合作用速率和籽粒的胚乳细胞数,饱满籽粒百分率以及千粒重显著增加。     

镉胁迫对不同水稻基因型植株生长和抗氧化酶系统的影响

 以籽粒镉积累水平不同的两种品种（丙97252，低积累型；秀水63，高积累型）为材料，研究了镉胁迫对水稻植株生长和抗氧化酶系统的影响。采用水培试验，镉处理设0.0、0.1、1.0和5.0 μmol/L 4个水平。结果表明，镉胁迫抑制植株生长和叶绿素合成，改变植株丙二醛（MDA）含量和超氧物歧化酶（SOD）、过氧化氢酶（CAT）、过氧化物酶（POD）活性。在抗氧化酶活性上，根和地上部对镉胁迫的反应存在着差异。总体上，SOD、CAT和POD活性随镉水平的提高而减少，而MDA含量则表现相反。根和地上部MDA含量随着培养液中镉浓度提高而增加，且增加幅度秀水63明显大于丙97252。与对照相比，生长在5.0 μmol/L Cd处理下的植株， SOD活性在孕穗期下降46%~52%，在分蘖期仅下降13%~19%。高浓度镉胁迫下，两品种在MDA含量的增加幅度和叶绿素含量的降低幅度上表现不同，显示出它们对镉的耐性存在着差异。     

盐胁迫条件下杂交水稻种子发芽特性和幼苗耐盐生理基础

 两个杂交稻组合汕优10号和两优培九种子分别放在H2O、50 mmol/L、100 mmol/L和150 mmol/L的NaCl溶液中于30℃下发芽，测定种子发芽性能和淀粉酶活性及幼苗保护酶活性、丙二醛（MDA）含量和脯氨酸、可溶性糖、果糖和蔗糖等相容性溶质含量。结果表明，盐胁迫条件下杂交水稻种子平均发芽时间延长，发芽指数降低，但发芽势和发芽率变化不明显。盐胁迫后明显降低两优培九种子α，β-淀粉酶活性，而汕优10号中， 除在50 mmol/L NaCl溶液中α-淀粉酶活性高于对照（H2O）外，其余处理均降低了α，β-淀粉酶活性。不同盐胁迫程度下杂交水稻幼苗超氧物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶(CAT)活性和丙二醛含量及脯氨酸、可溶性糖、果糖和蔗糖等相容性溶质含量变化有差异，但未见规律性趋势。杂交水稻幼苗相对含水量和耐盐比率随着盐胁迫程度加深而明显下降。试验还表明，盐胁迫条件下杂交水稻组合汕优10号种子发芽性能比两优培九好，淀粉酶和保护酶活性、相容性溶质含量和相对含水量及耐盐指数和耐盐比率也均高于两优培九，说明汕优10号幼苗耐盐性强于两优培九。     

蔗糖和激素对水稻离体培养穗籽粒生长的影响

 应用离体穗培养技术研究了蔗糖浓度和不同植物激素对不同源库类型水稻品种籽粒的溶液和蔗糖吸收以及干物积累的影响。离体培养穗的籽粒吸收溶液量随培养液蔗糖浓度的升高而增加，但高蔗糖浓度(盐粳2号为160 mmol／L，亚优2号和汕优63为320 mmol／L)时有抑制作用。离体培养穗对蔗糖的吸收随蔗糖浓度升高而增加。供试的不同源库类型品种籽粒干物质和淀粉的积累均受低蔗糖浓度促进，但进一步提高蔗糖浓度则抑制了盐梗2号和汕优63的籽粒增重和淀扮积累。 GA₃和玉米素对汕优63籽粒的溶液吸收和干物质积累有促进作用，GA₃ 对盐粳2号和玉米素对亚优2号籽粒干物质积累有促进效应，但溶液的吸收则有抑制作用。由此初步认为籽粒对水分和蔗糖吸收是两个既相互依赖又相互独立的过程，籽粒糖分的积累与同化物的供应及库强的调节有关。本文还对激素的可能作用进行了讨论。     

杂交稻旗叶衰老过程中超氧自由基与超氧物歧化酶活性的变化

植物叶片的衰老、光氧化与体内自由基的积累和膜脂过氧化有关。由于超氧阴离子的累积,导致叶绿素和蛋白质降解。而在植物体内也存在着清除自由基的酶系统(如SOD,CAT,POD等),对防止叶片衰老和强光等逆境的胁迫起着重要的保护作用。目前用不同水稻品种来研究叶片超氧自由基和内源清除剂之间的关系报道较少。本文研究了在自然条件和光氧化条件下,叶片衰老和光氧化过程中产生的超氧自由基(O_2~■)和超氧物歧化酶(sOD)活性的关系,为生理育种,防止水稻后期叶片的早衰及光氧化伤害提供依据和鉴定指标。     

Response of Glutathione and Glutathione S-transferase in Rice Seedlings Exposed to Cadmium Stress

A hydroponic culture experiment was done to investigate the effect of Cd stress on glutathione content(GSH)and glutathione S-transferase(GST,EC 2.5.1.18)activity in rice seedlings.The rice growth was severely inhibited when Cd level in the solution was higher than 10 mg/L.In rice shoots,GSH content and GST activity increased with the increasing Cd level,while in roots,GST was obviously inhibited by Cd treatments.Compared with shoots,the rice roots had higher GSH content and GST activity,indicating the ability of Cd detoxification was much higher in roots than in shoots.There was a significant correlation between Cd level and GSH content or GST activity,suggesting that both parameters may be used as biomarkers of Cd stress in rice.     

Exogenous Peroxidase Mitigates Cadmium Toxicity,Enhances Rhizobial Population and Lowers Root Knot Formation in Rice Seedlings

Soil cadmium (Cd) causes toxicity and oxidative stress, alters biochemical processes and rootknot formation in rice. Irrigation of exogenous peroxidase (POX) together with its co-substrate H2O2(POXRice + H2O2), is likely to have protective effect upon the biochemical and nodular changes in ricegrown in Cd-rich soil. Exposure to Cd concentration of 1.00 mg/L increased oxidative stress, loss of cellviability, electrolyte leakage and root knot formation, whereas it significantly lowered the chlorophyll leveland rhizobium growth in rice. Irrigation of exogenous POXRice + H2O2 to Cd-stressed rice seedlingsreversed the Cd-induced alterations in rice to levels similar in control (non-stressed) seedlings. Resultsprovided strong evidence of exogenous POXRice + H2O2-mediated reversal and restoration of physiologicaland biochemical processes as well as increased resistance of rice seedlings to root knot formation.Irrigation with POXRice + H2O2 appeared to contribute towards bringing normoxic conditions in the otherwisehypoxic soil environment by enhancing the O2 in pot-experiments due to reduced Cd uptake, enhancedmineral homeostasis of essential elements viz. P, Fe, Mo, Mg and Mn for maintenance of root architecturedamaged by lipid peroxidation and reduction in oxidative stress by reducing Cd-induced reactive oxygenspecies generation. Therefore, the mitigation of Cd-toxicity in rice through this novel approach appeared tobe a promising mode to limit Cd-uptake, modulate protective and tolerance mechanisms for sustainablerice yield in Cd-contaminated rice-croplands and prevent nematode attack in rice, however, more detailedstudies are needed prior to large scale applications.     

Anaerobically Induced Proteins in Rice Seedlings

Abstract

Seven-day-old rice seedlings ( Oryza sativa L. cv. Kinuhikari) were subjected to anaerobic stress and their proteins were analyzed using SDS-polyacrylamide gel electrophoresis. Anaerobic stress caused only minor changes in the pattern of proteins in the shoots, but disappearance of many protein bands in the roots. Three anaerobic stress proteins (ANPs ; 36-, 40- and 87-kD protein) were selectively induced in both roots and shoots of the seedlings, and 36-kD ANP was identified as the glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) by limited N-terminal amino acid sequencing. Activities of GAPDH in the shoots and roots were increased by the stress for over 24 h, and were 3.4- and 6.2-fold greater than those in non-stressed seedlings at 24 h, respectively. These results suggest that anaerobiosis induces the production of ANPs including GAPDH in the seedlings, which may allow the seedlings to survive under the stress condition.     

孕穗期水稻不同功能叶的发育蛋白质组学分析

 从蛋白质组学角度对杂交水稻汕优63孕穗期4个叶位功能叶片蛋白质组变化进行了研究,以揭示水稻孕穗期叶片蛋白质组的表达差异。4个不同时期功能叶片蛋白质经双向电泳得到分离,应用Imagemaster 2D Elite 5.0软件对所得到的双向电泳图谱进行分析比较,共得到差异表达蛋白质点23个,差异表达蛋白质点经质谱分析,有14个得到鉴定。对鉴定出的蛋白功能进行研究,结果表明:差异表达的蛋白质大部分(9个)为参与光合作用和呼吸作用的蛋白质;其次为与蛋白结构及功能调控相关的蛋白质(3个),表达量表现为随叶位上升而上调;其余为参与氨基酸代谢的蛋白质(2个),表达量在较低叶位叶片中无显著规律,在剑叶中的表达量显著上调。蛋白质是细胞功能执行者,受细胞功能或细胞环境变化的影响,因此,上述蛋白质发生变化的原因应与水稻不同叶位叶片生长发育特性有关。     

镉胁迫对水稻光合、叶绿素荧光特性和能量分配的影响

 以浙农952为材料，用水培法研究了不同浓度镉（Cd）胁迫对水稻光合、叶绿素荧光特性和能量分配的影响。结果表明，低浓度Cd（0.1 mmol/L）胁迫下水稻净光合速率（Pn）、Fv/Fm、ΦPSⅡ和qN均略有上升，qP无显著变化，这说明在0.1 mmol/L浓度的Cd胁迫下水稻充分启动了能量耗散途径，耗散了过剩光能，使光合机构免受伤害。高浓度Cd（1.0 mmol/L）处理后Pn 、Fv/Fm、ΦPSⅡ均显著降低，qP和qN也大幅下降，在能量分配方面D和Ex显著上升，P显著下降，这说明在1.0 mmol/L浓度的Cd胁迫下，水稻PSⅡ受到伤害，某些能量耗散途径受阻，过剩的激发能不能及时耗散又导致活性氧的产生，最终引起抗氧化物酶活性的升高和叶绿体结构的破坏。     

植物生长物质冠菌素诱导旱稻、水稻幼苗抗旱性的效应及生理机制

以旱稻 297 和水稻越富幼苗为材料,在 20% PEG 模拟干旱条件下,研究了植物生长物质冠菌素处理对叶片水分状况、质膜透性、渗透调节物质（脯氨酸、可溶性糖和可溶性蛋白）及内源激素（ABA、IAA 和GA3）含量的影响。干旱胁迫下,冠菌素处理可以维持旱稻 297（0.01 μmol/L）和越富（0.1 μmol/L）较高的叶片相对含水量,促进幼苗叶片中脯氨酸、可溶性糖和可溶性蛋白的积累,降低质膜透性,维持细胞质膜的完整性;同时,冠菌素（0.01 和0.1 μmol/L）处理明显促进旱稻 297 和越富幼苗叶片中ABA的积累,并改变了 IAA 和GA3 的浓度及比例。冠菌素处理能改善旱稻和水稻幼苗耐干旱胁迫的能力,最适浓度分别为0.01 μmol/L和0.1 μmol/L。     

外源水杨酸对杂交稻幼苗抗白叶枯病的效应

杂交水稻幼苗感染白叶枯病菌后,其叶片中ＳＯＤ和ＰＯＸ活性明显增强,ＭＤＡ含量和膜透性显著提高。经过一定浓度水杨酸（ＳＡ）浸种或３叶期幼苗根际预处理,ＳＯＤ和ＰＯＸ活性呈增长趋势,而膜脂过氧化作用受到不同程度的抑制。３叶期幼苗施用ＳＡ比浸种处理效果好。感病组合汕优６３的变化幅度大于抗病组合威优６号。可认为ＳＡ对杂交稻幼苗抗白叶枯病有一定效应。     

杂交稻旗叶衰老过程中超氧自由基与超氧物岐化酶活性的变化

The O₂^- production, scavenging capacity, photooxidation, protein content, and SOD activity were studied in leaves of Yayou 2 (indica-japonica hybrid rice) and Shanyou 63 (indica hybrid rice) at filling stage. It was shown that protein content decreased gradually with days of heading in two varieties in natural condition, and the decrease in Shanyou 63 was more than that in Yayou 2. The rate of O₂^- production and SOD activity increased from heading to 20 days after heading, then decreases but still maintained certain levels until 30 days after heading in two varieties. O₂^- production rate in Shanyou 63 was more than that of Yayou 2, but SOD activity was contrary in two varieties. In addition, the change of O₂^- and SOD under photooxidation condition was consistent with the tendency above under natural condition. It was also shown that there was negative correlation between protein degradation and O₂^- / SOD in the process of senescence in flag leaves after 20 days of heading. It is suggested that O₂^-/SOD could be used as an index of scavenging ability for O₂^- during senescence in rice.     

根际促生细菌对干旱胁迫下水稻生理特性的影响

【目的】本研究旨在探究植物根际促生菌蜡状芽孢杆菌F06菌株对不同干旱胁迫下水稻汕优63生理特性的影响。【方法】在盆栽试验条件下,以水稻汕优63为种植材料,研究了轻度(LD)、中度(MD)、重度(SD)3个干旱强度下接种蜡样芽孢杆菌(Bacillus cereus)F06对水稻生理特征的影响。【结果】与正常水分管理相比,干旱胁迫(DS)下水稻叶片光合速率(P_n)和气孔导度(g_s)逐渐降低;而干旱胁迫下接种F06可显著减缓P_n和g_s下降,与不接种(NP)处理相比,P_n和g_s分别增加7.67%、12.97%、18.14%和11.51%、16.63%、17.07%,且呈现出随着干旱胁迫程度的提高,增幅增大的趋势。干旱胁迫下水稻叶片初始荧光(F_o)、非荧光淬灭系数(NPQ)显著上升,最大光化学效率(F_v/F_m)、光化学猝灭系数(q_P)显著下降;而干旱胁迫下接种F06可显著抑制F_o、NPQ升高和F_v/F_m、q_P降低,明显改善水稻叶片光能转换效率。干旱胁迫下接种F06虽然不能改变叶片水势、相对含水量和相对电导率的变化趋势,但可以有效降低其变幅。正常水分处理下接种F06虽然没有增加光合色素含量,但干旱环境下显著抑制了光合色素的分解或降低。干旱显著降低了水稻叶片和根系细胞分裂素(CTK)含量,增加了叶片中脱落酸(ABA)的含量;在干旱胁迫下,接种F06可显著提高叶片和根系中CTK的含量。【结论】由此可见,干旱生境下接种F06,可调节植物体内的激素含量,减少干旱胁迫下光合色素的分解或流失,提高光合速率,增强水稻在干旱环境中的适应能力。     

水稻籽粒灌浆期间茎鞘贮存物质含量变化及其影响因素研究

 以库限制型水稻品种盐粳235和源限制型水稻品种汕优63和亚优2号进行研究，水稻籽粒的灌浆因品种源库关系而有很大差异，剪叶或疏花对强势粒的灌浆影响不大，但显著地延缓或加速弱势粒的灌浆进程，库限制型品种盐粳235抽穗前叶鞘和茎秆中临时性贮存物质积累高峰出现迟，单位库容占有贮存性物质比例低，两类型品种茎鞘中贮存物质均在籽粒发育早期输出，但盐粳135的输出持续时间长于汕优63和亚优2号的。疏花、剪叶和激素处理主要通过改变库源大小，增加或减少单位库容占有光合同化物比例及改变茎鞘贮存物质在整个籽粒灌浆物质中的比例来影响籽粒生长和发育。还就茎鞘贮存物质在籽粒早期发育中的作用进行了讨论。     

PEG 6000胁迫对小麦三叶期蛋白表达的影响

为了探索干旱胁迫下小麦蛋白质表达的变化,选取旱地品种烟D27、 陕优225和水地品种新麦18、小偃22、1031为材料,在小麦三叶期,用18%和35%两种浓度的聚乙二醇(PEG6000)模拟干旱胁迫,处理待试小麦24 h后,用TCA丙酮法提取叶片全蛋白,通过SDSPAGE电泳分析小麦在干旱胁迫后的蛋白质带谱,并运用高效液相色谱质谱联用技术鉴定了新麦18的35.0 kD干旱敏感蛋白条带。结果表明,PEG6000胁迫后分子量28.0 kD蛋白在水地品种中消失,在旱地品种中正常表达或诱导表达。此外,干旱胁迫使抗旱品种烟D27诱导产生33.0和23.0 kD干旱应答蛋白条带,导致抗旱性弱的水地品种新麦18的35.0 kD蛋白条带消失。对新麦18的 35.0 kD蛋白条带进行液相色谱分离结合质谱分析后发现其含有17种与基本生命活动有关的蛋白,这些蛋白与小麦对干旱胁迫的适应性代谢调节有关。