干旱胁迫对锦灯笼种子萌发及幼苗生理特性的影响

以锦灯笼种子为试材,用3%、5%、8%和10%4个浓度的PEG-6000溶液模拟不同的干旱胁迫条件,研究锦灯笼种子萌发情况和幼苗生理特性,以期了解其抗旱性。结果表明:1)随着PEG-6000浓度的加大,锦灯笼种子的发芽率和发芽势先增加后减少,浓度为5%时最大,株高也是在胁迫浓度为5%时达到最高,这说明一定程度的干旱胁迫可以促进种子萌发及幼苗生长。2)丙二醛含量、可溶性糖含量和电导率随着PEG-6000浓度的增加表现为持续上升的趋势,过氧化物酶(POD)活性则呈现先上升后降低的趋势。     

外源NO及反向调控PEG胁迫下苜蓿萌发种子抗氧化酶及其同工酶动态的研究

以紫花苜蓿为材料,用一氧化氮供体硝普钠(SNP)及一氧化氮清除剂c-PTIO对苜蓿种子进行浸种处理,采用分光光度法和同工酶电泳技术来研究外源NO及反向调控对PEG胁迫下紫花苜蓿抗氧化酶活性及其同工酶的影响,并探讨NO调控苜蓿种子耐旱性的生理机制。结果表明:PEG胁迫下外施0.1 mmol·L^-1 SNP,第6天时较PEG处理POD活性降低了32.72%;第4天时SOD、CAT活性较PEG处理升高了10.48%、23.60%,有效缓解了PEG对紫花苜蓿萌发中种子的氧化损伤,提高其抗氧化能力。PEG胁迫下添加 c-PTIO抑制了苜蓿萌发期的抗氧化系统活性。从同工酶的谱带数量和强弱来看,POD同工酶各区带活力均随PEG胁迫时间的延长而增加,在第2天酶带只有1条,而第4天酶带呈现9条;SOD和CAT同工酶表达量变化不显著,但酶带强弱有一定变化,S3酶带随处理时间的延长表达量逐渐减弱;CAT同工酶谱带则一直保持2条带,无明显强弱变化。因此,外源NO在PEG胁迫下紫花苜蓿萌发中抗氧化酶快速响应并在维护氧自由基代谢平衡中起重要保护作用。     

渗透胁迫下紫花苜蓿生长及ABA受体基因的表达

以紫花苜蓿不同品种(‘三得利'‘新牧2'‘赛迪5'‘WL656'‘WL168'‘WL903'‘WL712'‘WL343')为试验材料,研究150g/L PEG-6000胁迫下,幼苗抗旱性与ABA受体基因表达的关系。结果表明,150g/L PEG处理下,不同苜蓿幼苗较对照根系缩短(10.58%~15.88%)、苗长减少(18.06%~23.71%)、鲜干质量比降低(24.51%~37.81%)、相对含水量下降(13.31%~16.71%),差异显著;脯氨酸质量分数均显著升高(21.24%~39.04%),抗旱性强弱为:‘WL168'‘新牧2'‘WL712'‘赛迪5'‘WL343'‘WL656'‘WL903'‘三得利';‘WL168'的ABA受体Medtr1g016480和Medtr5g083270基因的表达量显著升高,二者在根茎叶中的表达量分别为6.10、1.79、3.04和6.51、1.67、1.89,其表达量强弱为:根叶茎。渗透胁迫下,苜蓿ABA受体Medtr1g016480和Medtr5g083270基因的表达量显著升高,对苜蓿生理生化反应及其生长起到一定的调节作用,进而影响苜蓿的抗旱能力。     

柱花草叶肉原生质体提取与瞬时表达体系构建

柱花草(Stylosanthes guianensis)是一种重要的热区豆科牧草,为利用原生质体瞬时表达体系开展柱花草基因功能研究,以20 d龄期‘热研5号’柱花草子叶为试验材料,通过正交试验考察不同因素对原生质体分离及转化效率的影响。结果表明：采用1.5%纤维素酶、1.25%离析酶、0.5 mol·L^-1甘露醇和4 mmol·L^-1MES酶解液组合,酶解时间为16 h时,原生质体的产量和活性均达到最佳,分别为(4.567×10⁶)个·g^-1和92.271%;原生质体浓度为(6×10⁵)个·mL^-1、质粒浓度为0.6μg·μL^-1,PEG-4000浓度为50%、转化时间为5 min时,转化效率最高,可达52.524%。构建了目标蛋白SgRKL1表达载体pA7-BIUTNT::SgRKL1-GFP,利用PEG介导法将其转入柱花草原生质体,激光扫描共聚焦显微镜检测结果显示SgRKL1蛋白定位于细胞膜上,说明所建立的柱花草叶肉原生质体瞬时表...     

两种卵黄抗体不同提取方法的比较试验

卵黄抗体(Ig Y)具有许多其他类型抗体所没有的优点,其分离纯化的方法有有机物抽提法、盐析法、水稀释法等。试验分别用聚乙二醇(PEG)6000和氯仿来提取卵黄抗体,对其收集的抗体量和抗体效价进行比较分析。结果表明:100 g的卵黄用PEG6000法比氯仿法多收集71 m L卵黄抗体溶液,且PEG6000法的成本远远低于氯仿法,有很好的经济效益,而PEG6000法提取的卵黄抗体效价比氯仿法略低一个滴度。     

PEG处理对柠条种子发芽的影响

通过选用渗透调节剂聚乙二醇(PEG)处理柠条种子,并对柠条种子的发芽情况进行试验研究,结果表明:在水分胁迫条件下,柠条种子萌发存在一定的差异;一定浓度范围内,随着PEG溶液浓度的增强,柠条种子的发芽率提高,发芽速度加快。据文献报道,当PEG处理的强度达到一定水平时,会对种子的萌发产生抑制作用,但本试验还没有得到对柠条种子产生抑制作用的PEG浓度。     

Seed germination ofThuja orientalis under PEG osmose treatment

Abstrct Thuja orientalis seeds was treated by different molecular weight, different concentration and different lasting time in the Seedling Laboratory of Northeast Forestry University. The results are as follows:(1) it shows significant promoting effects on the germination viability of seeds by different molecular weight of PEG(-6000, -10000 and -20000), and PEG-10000 is the best. (2)Different effects on germination viability, germination rate and simple vigor index are made by different concentration of the same weight molecular(PEG-10000), the promoting effects are shown when the concentration is below 30%, and from 20% to 30% is the best range, but inhibitions will be shown if the concentration is more than 35%. (3)The most suitable lasting time of osmose treatment of PEG-10000 with 20% concentration is from 2 to 4 days, not more than 6 days, but it will show significant inhibition if the treated time is more than 8 days. (4)The treatments of PEG-10000 and PEG-20000 with higher molecular weight can increase the germination rate and seed vigor, and decrease the electric conductivity of the seed soaking liquid after the natural air-dry treatment. The suitable time of natural air-dry is about 6 to 8 hours. It is also concluded that the PEG osmose treatment is an effective method to increase the germination rate by the field simulation in the laboratroy.     

Changes of plasma membrane H<Superscript>+</Superscript>-ATPase activities of <Emphasis Type="Italic">Glycine max</Emphasis> seeds by PEG treatment

The soybean （Glycine max） Heihe No. 23 is sensitive to imbibitional chilling injury. Polyethylene glycol （PEG） treatment can improve chilling tolerance of soybean seeds to a certain extent. The changes of hydrolytic ATPase in plasma membranes and H^＋-pumping responses in soybean seeds were investigated during PEG treatments. Effects of exogenous calcium and exogenous ABA on the hydrolytic ATPase were also examined in order to understand the mechanism of chilling resistance. Highly purified plasma membranes were isolated by 6.0% aqueous two-phase partitioning from soybean seeds, as judged by the sensitivity of hydrolytic ATPase to sodium vanadate. PEG treatment resulted in a slight increase of the hydrolytic ATPase activity in 12 h. Then the activity decreased gradually, but still higher than the control. The H^＋-pumping activity increased steadily during PEG treatment. Exogenous calcium had both activating and inhibiting effects on the hydrolytic ATPase, but the activity was inhibited in soybean seeds treated with exogenous ABA. Results suggested that PEG treatment, not the exogenous calcium and ABA, up-regulated H^＋-ATPase activities in soybean seeds.     

PEG-6000胁迫下3种外生菌根真菌的生长特性

为了解不同外生菌根真菌的抗旱能力及机制，筛选具有较强抗旱性的优良菌株。选取琥珀乳牛肝菌(sp)、粘盖乳牛肝菌(sb)、橙黄硬皮马勃(sc)3种外生菌根真菌，采用液体培养和固体培养2种方式，利用PEG-6000模拟干旱条件，研究不同干旱胁迫程度对3种外生菌根真菌生长及生理变化的影响。结果表明，在不同PEG-6000浓度下,sb菌丝生长速率一致，sp在0~21 d的菌丝生长速率较高，sc则在14~28 d才呈现速生期；轻度干旱胁迫（PEG-6000浓度为10％~20％）有利于sb和sp生物量的积累，增加sb对H⁺的分泌，并提高sb的SOD活性，降低sb的MDA含量，而sc的生长则受到PEG-6000的抑制；在PEG-6000处理下，3种外生菌根真菌的菌丝形态出现了不同程度损伤，随PEG-6000浓度升高菌丝形态损伤进一步加重，但sb菌丝形态结构完整度高于sp与sc。因此，轻度干旱胁迫利于sb的生长，3种外生菌根真菌抗旱性强弱为:sb>sp>sc。本研究结果可为外生菌根真菌抗旱机制的深入研究及抗旱菌种的筛选提供参考。     

大豆种子发芽期耐旱性鉴定的适宜PEG-6000浓度筛选

以抗旱性不同的3个大豆品种(湘豆3号、Lee68和宁镇1号)为试材,研究了不同浓度(5%、10%、15%、20%、25%和30%)的PEG-6000模拟发芽期干旱胁迫对大豆种子发芽、子叶苗膜脂过氧化以及渗透调节物质含量的影响。结果表明:10%以下的PEG-6000能起引发作用,促进大豆种子的萌发,显著提高发芽势和发芽率;随着PEG-6000浓度的升高,大豆种子的发芽明显受抑制,当PEG-6000浓度大于25%时抑制更明显,发芽势、发芽率、发芽指数和简易活力指数显著下降;随着PEG-6000浓度的升高,大豆子叶苗中的丙二醛(MDA)、脯氨酸(Pro)、可溶蛋白和可溶性糖含量显著增加,PEG-6000浓度在15%~25%间增加幅度最大,PEG-6000浓度小于15%或大于25%时增加不明显。综合种子发芽、子叶苗的膜脂过氧化程度以及渗透调节物质含量的变化,可初步确定室内采用PEG-6000浸种法模拟干旱胁迫鉴定大豆发芽期耐旱性的适宜浓度为20%~25%。