滇重楼种子内源抑制物质的活性研究

以滇重楼种皮和胚乳为材料,用甲醇浸提得到种皮浸提液和胚乳浸提液,配制成浓度梯度为0.02、0.04、0.06、0.08g/mL的处理液,以蒸馏水为对照,研究不同浓度种皮和胚乳浸提液对受体植物白菜(油脂类)、小麦(淀粉类)、绿豆(蛋白类)种子发芽率、幼苗苗高和根长、抗氧化酶活性的影响,探讨滇重楼种子内源抑制物质的特性,以揭示滇重楼种子休眠的生理机制。结果表明:(1)滇重楼种皮和胚乳中均含有抑制种子萌发与幼苗生长的物质,且抑制作用大小表现为胚乳大于种皮。(2)3种受体植物幼苗对滇重楼种皮、胚乳甲醇浸提液的敏感性大小为白菜(油脂类)显著高于绿豆(蛋白类)和小麦(淀粉类)。(3)滇重楼种皮、胚乳甲醇浸提液对种子萌发及幼苗生长各项指标的抑制作用强度依次为:发芽率幼苗根长幼苗苗高鲜重。(4)滇重楼种皮、胚乳中的抑制物质均能够使3种受体植物幼苗产生生理响应,启动抗氧化酶体系,其中:随着滇重楼种皮甲醇浸提液浓度的升高,受体幼苗SOD与POD活性先升后降,CAT活性持续下降;胚乳甲醇浸提液处理后,受体幼苗SOD和POD活性较对照均显著增强,而CAT活性显著下降。研究认为,滇重楼种皮及胚乳甲醇浸提液均能够抑制受体植物种子萌发及幼苗生长,影响幼苗抗氧化酶活性,但滇重楼种皮与胚乳中所含抑制物在活性上存在差异。     

桃儿七种子休眠解除过程中细胞壁代谢及种皮超微结构的变化北大核心CSCD

为探究低温层积过程中桃儿七种子细胞壁代谢及种皮超微结构与休眠解除的内在联系,该研究通过低温层积解除桃儿七种子休眠,分析休眠解除过程中种子不同部位细胞壁组分及相关代谢酶的变化,同时利用扫描电镜对种皮的超微结构进行观察。结果表明,(1)桃儿七种皮主要由角质层、栅状石细胞层及海绵组织层3层构成,在层积过程中,种皮内部的海绵组织逐步疏松膨胀,种皮表面破损加剧;(2)种子不同部位的细胞壁组分具有明显差异,整个层积过程中,种胚、种皮和胚乳中的纤维素含量均在层积中期(45 d和60 d)降至最低,3个部位的纤维素酶活性在层积中期对应升高;种胚和种皮内的半纤维素含量均在层积中期显著下降,种皮中甘露聚糖酶活性和木糖苷酶活性在层积中期时相应达到最大;3个部位的果胶含量均在层积后期(75 d和90 d)时显著下降,而种皮和胚乳中多聚半乳糖醛缩酶活性也在层积后期相应升高;(3)种胚和胚乳内过氧化物酶活性在层积75 d和90 d时明显下降,而SOD活性在此时显著上升。(4)种子不同部位3种木质素单体的组成比例具有明显区别,同时3种木质素单体含量均随层积时间的延长而显著降低,且胚乳和种皮中的S-木质素含量对种子萌发存在显著的负向影响关系。研究认为,在低温层积过程中,桃儿七种子内细胞壁组分纤维素、半纤维素及木质素的逐步酶解,活性氧作用下的细胞壁松弛以及海绵组织层的疏松膨胀和种皮的破裂,破坏了细胞壁的刚性结构,促使种子机械束缚力降低,吸水性能提高、胚根生长能力增强,最终导致其休眠解除。     

外源水杨酸对NaCl胁迫下大豆种子萌发和幼苗生长生理的影响

以大豆种子、幼苗为试验材料,采用砂培的方法,研究了0.2mmol·L-1外源水杨酸(SA)对100mmol·L-1 NaCl胁迫下大豆种子萌发、幼苗形态及生物量、膜脂过氧化和抗氧化酶活性的影响。结果显示:NaCl胁迫下,大豆种子萌发和幼苗生长受到显著抑制,且随着胁迫时间的延长(0~3d),大豆幼苗相对电解质渗漏率、硫代巴比妥酸活性产物(TBARS)含量显著升高,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性均明显降低。外源SA促进NaCl胁迫下大豆种子萌发和根茎生长,增加幼苗生物量积累,降低幼苗叶片相对电解质渗漏率和TBARS含量,增强其叶片SOD、CAT、APX活性。研究表明,NaCl胁迫能显著抑制大豆种子萌发和幼苗生长,而一定浓度的外源SA能有效提高NaCl胁迫下大豆种子活力及幼苗抗氧化酶活性,减轻膜脂过氧化程度,缓解NaCl胁迫所造成的伤害,提高大豆幼苗抗盐胁迫的能力。     

柃属植物种子微形态特征及其系统学意义

利用扫描电镜对35种1变种柃属(Eurya)植物种子的微形态特征进行了研究,并采用相对大小(长×宽)RS值和种皮表面纹饰作为种子形态比较的特征参数。结果显示,柃属种子根据相对大小RS值可划分为小型(RS<1.0),中型(1.03.0)三个等级,最小的种子相对大小不足0.5,最大的种子相对大小超过10;种皮表面纹饰分为负网纹和网纹两种类型,网纹类型在网眼的方向性、网眼形状、网眼深度、网脊特点等方面呈现出种间多样性,可用于类群的比较和鉴别。文中同时还探讨了种子大小及种皮微形态在属内的系统发育关系,推测了种皮表面纹饰的演化路线,研究结果表明种子的微形态特征对本属植物系统分类及进化研究具有重要意义,可为属内一些关键种系统位置的分析提供重要佐证。     

大豆种子超氧物歧化酶的研究

大豆种子的SOD能抑制肾上腺素的自动氧化和氮蓝四唑(NBT)的光化还原。可以根据SOD对NBT光化还原的抑制作为酶活性的定量测定。大豆SOD活性被KCN抑制,但不受氯仿-乙醇影响,表明了大豆SOD为Cu-Zn SOD。大豆SOD具有一定的耐热性,它的粗提液经聚丙烯酰胺凝胶电泳及NBT的酶活性染色,在蓝色背景上有5～6条无色透明的同工酶谱带。大豆SOD中性粗提液经纯化,比活提高13倍。纯化的大豆SOD和牛血SOD有同样的光吸收波谱。     

四棱豆不同节位各器官中SOD活性的测定

测定实验表明 :位于四棱豆植株中部节位叶片中的SOD活性高于植株下部和上部节位叶片中的SOD活性 ;而位于下部节位豆荚和种子中的SOD活性要高于位于上部节位豆荚和种子中的SOD活性 ,且随着结荚部位升高 ,SOD活性呈降低趋势。     

生理成熟期高温高湿胁迫对春大豆种子活力、主要营养成分及种皮结构的影响

采用盆栽方式,研究了宁镇1号和湘豆3号2个春大豆品种在种子生理成熟期受到高温高湿胁迫[(40±2)℃/(30±2)℃、相对湿度(95±5)%/(70±5)%、10 h/14 h(昼/夜)]后种子活力指标、主要营养成分及种皮解剖结构的变化.结果表明:生理成熟期植株受到高温高湿胁迫后,2个春大豆品种种子生活力、发芽势、发芽率以及脱氢酶、酸性磷酸酶活性呈下降趋势,种子细胞膜透性增大,浸种液中的H+、可溶性糖和亮氨酸含量上升,种子中的脂肪和蛋白含量无规律性变化,种皮与种脐解剖结构发生改变;短期的高温高湿胁迫(5 h)对春大豆种子活力的影响不明显,胁迫时间超过48 h,种子活力相关指标急剧降低;湘豆3号较耐高温高湿胁迫,表现为发芽势和相关酶活性下降及浸种液外渗物含量增加的幅度小,种皮致密、厚实,种脐结构完整等特征.     

石竹属种子形态及其分类学意义

利用扫描电镜对石竹属(Dianthus)10种植物的种子宏观形态和种皮的微观形态特征进行观察,首次对该属的种皮微观形态特征进行系统报道。研究结果表明:该属的种子宏观形态可分为3种类型:椭圆形、卵圆形和三角形。种皮表面微观形态均为条形—负网状纹饰或负网状纹饰,按网眼形状将其分为了3个类型,种间网眼大小,网脊宽度具明显差异,这对于石竹属的系统分类具有一定的参考价值。     

大豆卵磷脂对小鼠的抗疲劳和抗氧化作用研究

目的:研究大豆卵磷脂的抗疲劳及抗氧化作用。方法:小鼠经口给予大豆卵磷脂30天后,采用负重游泳实验,观察记录小鼠游泳死亡时间;检测血清尿素氮、肝糖原;测定血清和肝匀浆超氧化物歧化酶(SOD)活性、谷胱甘肽过氧化物酶(GSH-Px)活力、丙二醛(MDA)含量。结果:给予大豆卵磷脂后,与对照组相比,实验组小鼠负重游泳时间明显延长,肝糖原消耗量减少,降低运动后血清尿素氮水平(P<0.05);升高小鼠血清和肝匀浆SOD活性及GSH-Px活力,降低MDA的含量(P<0.05)。结论:大豆卵磷脂具有抗疲劳和抗氧化作用。     

花生种皮抗黄曲霉差异基因表达分析

选择高抗黄曲霉花生种质J11与高感种质金花1012为研究材料,利用基因芯片技术开展了抗病差异基因表达分析研究。试验共使用61078组大豆杂交探针。初步结果表明,与金花1012相比,J11种皮共有417个差异基因表达量上升,650个差异基因表达量下降,其中两份种质的种皮过氧化物酶（POD）差异基因表达量存在明显差异。为初步验证基因芯片分析结果,进行了POD活性测定。结果表明,种子接近成熟前,J11种皮的POD活性迅速上升,明显高于金花1012水平;金花1012则变化不明显,与相关基因差异表达量基本一致。     

Study on the Superoxide Dismutase of Soybean Seeds from Different Species in Subgenus

The SOD isoenzymes of different soybeans were studied in this experiment and the results showed that: 1. there were 7 SOD isoenzyme bands in all soybean genotypes used in this experiment, 2. the SOD activity (unit/g fr. wt.) in different soybean seeds decreased as the evolutionary process advanced, 3 the SOD activity decreased as the wt./100 seeds increased, and the color of seed coat changed from black to yellow. The relationship between SOD activity and the evolutionary process was discussed.     

PEG处理大豆种子对下胚轴脂氧合酶活性及脯氨酸含量的影响

选用三种不同浓度的PEG 6000处理抗旱性不同的两个大豆品种“庆选101”和“哈85-6439”。研究结果表明:PEG处理大豆种子可以显著地降低下胚轴中的LOX活性,pro含量也明显下降,而蛋白质的含量却迅速上升。这与水分协迫条件下植物体内大量积累pro的试验结果相反,并且pro含量的下降与LOX活性的降低、蛋白质含量的升高密切相关,同时种子的发芽率明显提高。     

大豆出苗期和苗期对盐胁迫的响应及耐盐指标评价

比较了4个大豆品种出苗期和苗期的耐盐性,测定150 mmol/L NaCl胁迫下的株高、下胚轴长、侧根数、地上干/鲜重、根干/鲜重、MDA含量、SOD活性、游离Pro含量,并将幼苗移栽到田间生长至成熟。结果表明:出苗期和苗期盐胁迫下4个品种的株高都显著降低、地上干/鲜重和根干/鲜重降低;出苗期胁迫侧根数减少,下胚轴长降低;而苗期胁迫侧根数增加,下胚轴长升高。未胁迫条件下,出苗期和苗期耐盐性强的品种22021-1的MDA含量和SOD活性高于耐盐性弱的品种22293-1。胁迫后,22021-1的MDA含量降低、SOD活性升高,其MDA含量分别比对照低51.03%和21.45%,SOD活性比对照高5.85%和45.77%;22293-1的MDA含量出苗期比对照高58.97%,苗期基本无变化,SOD活性出苗期和苗期升高都不显著。MDA和SOD可以作为大豆耐盐性筛选指标。早期的短时胁迫对不同耐盐性大豆品种的经济产量影响不同。     

3种进化类型大豆叶片的某些生理特性比较

半野生、半栽培和栽培大豆叶片可溶性糖和可溶性蛋白含量以及过氧化物酶（POD）和超氧化物歧化酶（SOD）活性均表现为随品种的进化而增加的变化趋势。POD活性在生育期中呈上升变化,SOD活性则随着生育进程逐渐下降。栽培大豆的可溶性糖含量和POD活性以上部叶片最高,半栽培大豆与栽培大豆类似,半野生大豆的变化则不明显。     

Species and Organ Diversity in the Effects of Hydrogen Peroxide on Superoxide Dismutase Activity In Vitro

Superoxlde dlsmutase （SOD） is ubiquitous in aerobic organisms and constitutes the first link In the enzyme scavenging system of reactive oxygen species. In the present study, species and organ diversity of SOD activity In a solution and In an in-gel assay system, as well as the effects of hydrogen peroxide （H202） on SOD activity, were Investigated. In a solution assay system, SOD activity of jackfruIt root, shoot, leaves, axes, and cotyledons, of maize embryos and endosperms, of mung bean leaves and seeds, of sacred lotus axes and cotyledons, and of rice and wheat leaves was Increased by 1-15 mmol/L H2O2. However, SOD activity In rice root and seeds, maize roots and leaves, mung bean roots and shoots, and wheat seeds was decreased by 1-15 mmol/L H2O2. The SOD activity of wheat root and soybean roots, leaves, axes, and cotyledons was Increased by 1-4 mmol/L H2O2, but was decreased by concentrations of H2O2 〉4 mmol/L. The SOD activity of soybean shoots was not affected by 1-15 mmol/L H2O2. The SOD activity In crude mltochondrla of jackfruIt, maize, and upas seeds, as well as In purified mitochondria of jackfruIt, was also Increased by 1-15 mmol/L H2O2. In the In-gel assay system, the SOD In jackfruIt cotyledons was comprised of Mn-SOD, Cu/Zn-SOD, and Fe-SOD, the crude mltochondria of jackfruit seeds and maizes embryo was comprised of Mn-SOD and Cu/ Zn-SOD, and the crude mltochondria of maize seeds was comprised of Mn-SOD only. In the present study, H2O2 markedly Inhibited Cu/Zn-SOD and Fe-SOD activity.     

1,2,4-三氯苯胁迫对萌发大豆种子中活性氧的影响

以盆栽法研究了不同浓度 1,2 ,4 三氯苯 (TCB)胁迫对萌发大豆种子中活性氧代谢的影响 .结果表明 ,10 0～ 30 0 μg·g-1TCB胁迫初期 (1～ 3天 )促使萌发大豆种子呼吸强度升高及其峰值提前出现 ,超氧阴离子自由基 (O2 - )及过氧化氢 (H2 O2 )的积累显著增加 ,同时伴随丙二醛 (MDA)含量升高 ,显示发生膜脂质过氧化作用 .TCB胁迫 1～ 6天使活性氧清除酶功能紊乱 ,其中过氧化物酶 (POD)活性升高 ,超氧化物岐化酶 (SOD)活性开始上升后转为下降 .在萌发大豆种子受TCB胁迫伤害过程中 ,活性氧代谢失衡造成的膜脂质过氧化将起着重要作用 .     

大豆PM34基因生物信息学预测及表达分析

以大豆叶片总RNA为模板,采用RT-PCR法从吉豆2号品种中克隆获得大豆种子成熟蛋白( PM34)基因序列,利用生物信息学方法对大豆PM34基因编码的蛋白进行预测。结果表明：该基因编码蛋白理论分子量为31.7 kDa,等电点为6.60,为亲水性蛋白;该蛋白中无跨膜结构;该蛋白中不存在信号肽。 PM34基因编码蛋白的二级结构中α螺旋占12.97％,无规则卷曲占41.30％,β折叠占45.73％。 PM34基因编码蛋白的三级结构预测表明,同源模建的模板是3ijr.1.A,是一种短链脱氢酶/还原酶,与该蛋白的同源性为54.65％。在进化关系上,与绿豆、苜蓿的亲缘关系相对较近。采用实时定量PCR方法( qRT-PCR),检测大豆PM34基因在大豆各器官中的表达方式,结果表明该基因在大豆根、茎、叶、花中的表达活性低,而在种子中,尤其是成熟种子中的相对表达活性很高。该研究结果为大豆PM34基因结构和功能的进一步研究奠定了基础。     

Enhanced levels of nicotianamine promote iron accumulation and tolerance to calcareous soil in soybean

Iron (Fe) is an essential nutrient in both plants and humans. Fe deficiency on calcareous soil with low Fe availability is a major agricultural problem. Nicotianamine (NA) is one of the Fe chelator in plants, which is involved in metal translocation into seeds, and serves as an antihypertensive substance in humans. In this study, soybean plants overexpressing the barley NA synthase 1 (HvNAS1) gene driven by the constitutive CaMV 35S promoter were produced using Agrobacterium-mediated transformation. The transgenic soybean showed no growth defect and grew normally. The NA content of transgenic soybean seeds was up to four-fold greater than that of non-transgenic (NT) soybean seeds. The level of HvNAS1 expression was positively correlated with the amount of NA, and a high concentration of NA was maintained in the seeds in succeeding generations. The Fe concentration was approximately two-fold greater in transgenic soybean seeds than in NT soybean seeds. Furthermore, the transgenic soybeans showed tolerance to low Fe availability in calcareous soil. Our results suggested that increasing the NA content in soybean seeds by the overexpression of HvNAS1 offers potential benefits for both human health and agricultural productivity.     

Phospholipid and triacylglycerol profiles modified by PLD suppression in soybean seed

Phospholipase D (PLD) is capable of hydrolyzing membrane phospholipids, producing phosphatidic acid. To alter phospholipid profiles in soybean seed, we attenuated PLD enzyme activity by an RNA interference construct using the partial sequence from a soybean PLDα gene. Two transgenic soybean lines were established by particle inflow gun (PIG) bombardment by co‐bombarding with pSPLDi and pHG1 vectors. The lines were evaluated for the presence and expression of transgenes thoroughly through the T₄ generation. PLD‐suppressed soybean lines were characterized by decreased PLDα enzyme activity and decreased PLDα protein both during seed development and in mature seeds. There was no change in total phospholipid amount; however, the PLD‐attenuated transgenic soybean seed had higher levels of di18 : 2 (dilinoleoyl)‐phosphatidylcholine (PC) and ‐phosphatidylethanolamine (PE) in seeds than the non‐transgenic lines. The increased polyunsaturation was at the expense of PC and PE species containing monounsaturated or saturated fatty acids. In addition to increased unsaturation in the phospholipids, there was a decrease in unsaturation of the triacylglycerol (TAG) fraction of the soybean seeds. Considering recent evidence for the notion that desaturation of fatty acids occurs in the PC fraction and that the PC → DAG (diacylglycerol) → TAG pathway is the major route of TAG biosynthesis in developing soybean seed, the current data suggest that PLDα suppression slows the conversion of PC to TAG. This would be consistent with PLD playing a positive role in that conversion. The data indicate that soybean PLD attenuation is a potentially useful approach to altering properties of edible and industrial soybean lecithin.     

沙芥种子发育过程中的脱水耐性

以发育过程中经脱水和未脱水处理的沙芥种子为试验材料,测定了其含水量、萌发率和抗氧化酶系统,探讨了沙芥种子脱水耐性与抗氧化系统之间的关系。结果表明：在20～60DAF,沙芥种子含水量逐渐下降,干重逐渐增加;60DAF种子具有萌发能力,萌发率为24％;且脱水可促进沙芥种子的萌发,人工脱水至含水量为12％和5％时,萌发率分别为56％和44％,自然脱水至含水量为12％和5％时,萌发率分别为52％和60％;发育过程中沙芥种子SOD活性逐渐降低,而CAT、POD、LOX活性以及MDA含量均呈上升趋势;在脱水过程中,随着种子含水量的下降SOD活性逐渐降低,CAT和LOX活性逐渐升高,而POD活性呈先降低后升高的变化趋势。脱水后,种子中MDA含量均高于CK。60DAF的沙芥种子已获得脱水耐性。