UV—B辐射增强对苹果采后炭疽病发病情况和抗病相关酶活性的影响

模拟UV-B增强的实验条件,在人工接种的情况下,比较了苹果采后炭疽病的发病情况,测定了一些与抗病相关酶的活性.UV-B增强后,苹果炭疽病斑直径、病斑扩展速率降低,发病指数较对照下降了58.83%.POD活性在处理后第7天时达最大值,为处理前的1.76倍,PPO活性在处理后第6天增加到对照的1.5倍.PAL活性在第6d提高1.74倍,几丁质酶和β-1,3葡聚糖酶活性都有所升高,且它们的变化时程一致.结果表明:UV-B增加限制了苹果采后炭疽病的发展,诱导果实中防御酶活性.     

果实抗病防御应答参与SO_2对‘红提’葡萄的采后保鲜

以本地产‘红提’葡萄果实为实验材料,运用SO_2两段释放处理结合冷藏的商业贮藏保鲜技术,研究保鲜剂SO_2对鲜食葡萄果实采后抵御病菌感染的作用机制.结果发现:使用SO_2保鲜剂可使‘红提’葡萄的贮藏期延长,在60 d贮藏期内SO_2处理组葡萄果实好果率明显高于对照组.贮藏期间,葡萄果皮中次生代谢途径的关键酶苯丙氨酸解氨酶活性升高,次生代谢产物总酚、类黄酮和木质素含量升高,其中SO_2处理组各指标的增幅明显高于对照组;多酚氧化酶活性在SO_2处理组和对照组中均降低,但SO_2组酶活性高于对照组,表明SO_2保鲜剂能够增强‘红提’葡萄果实次生代谢途径,促进次生代谢产物积累,提高细胞抗氧化能力和细胞壁结构防御能力.SO_2处理组葡萄果皮中几丁质酶和β-1,3-葡聚糖酶活性均显著高于同期对照组,说明SO_2处理能增强果实的抗病防御应答,提高果实抗病防御能力.本研究表明SO_2能介导‘红提’葡萄采后贮藏期间抗病防御应答,促进果实保鲜,延长果实采后贮藏期.     

有机溶剂和变性剂对枯草芽胞杆菌溶栓酶活性的影响

以枯草芽胞杆菌LD-8547菌株发酵液为材料,提取分离溶栓酶,研究了几种有机溶剂及变性剂对枯草杆菌溶栓酶活性的影响.结果表明,甲醇对溶栓酶表现为抑制作用,在10%-15%浓度范围时,其抑制作用最强;乙醇在5%～15%浓度范围内对酶活有激活作用,但随浓度的增大则转变为较强的抑制作用;正丁醇和异丙醇对酶活力则表现出强烈的抑制作用;丙三醇对酶活力有轻微的抑制作用,随着浓度增加抑制作用逐渐增强;异戊醇在5%～15%浓度范围内有一定的激活作用,但当浓度大于15%时则表现为抑制作用.醛类对酶活也有较强的抑制作用;丙酮、DMSO、DMF、SDS和异氰硫酸胍对酶活均有不同程度的抑制作用;脲浓度＜0.01 mmol/L时略有激活作用,随浓度增大转变为抑制作用.实验同时对EDTA在不同浓度和不同时间内对酶活力的影响进行了研究,结果表明,浓度＜2mmol/L的EDTA基本不影响酶活力,但随着浓度的增大,其抑制作用明显增大.本实验还以牛血为材料,对酶液进行了体外抗血凝和溶血栓实验,结果表明,该酶具有抗血凝和溶血栓效果.     

香蕉MaMPK1基因的克隆与表达模式

为研究香蕉MAPK1的序列特征及其在不同激素处理、逆境胁迫下的表达趋势,以‘天宝蕉’为材料,采用RTPCR技术克隆MaMPK1并对其进行生物信息学分析和不同处理下的表达模式分析.结果显示该基因编码区长为1182bp,可编码393个氨基酸.其编码蛋白具有STKc_TEY_MAPK结构域,属于MAPK基因家族TEY亚型A亚家族,是不稳定的脂溶性亲水酸性蛋白,无信号肽和跨膜结构,有多个磷酸化位点.亚细胞定位预测结果显示MaMPK1主要定位于细胞核.蛋白互作预测结果显示该蛋白与HSFA4A存在互作,暗示其可能在香蕉抗热反应过程中发挥作用.启动子顺式作用元件预测结果显示MaMPK1启动子包含多种激素和逆境胁迫相关作用元件.定量分析结果显示MaMPK1的表达受SA、45℃、低温和盐胁迫抑制,受茉莉酸甲酯(MeJA)和枯萎病菌侵染诱导上调,在脱落酸(ABA)处理后期极显著上调表达.本研究表明MaMPK1广泛参与香蕉逆境胁迫应答.(图9表1参35)     

稀土元素对怀槐悬浮培养细胞异黄酮合成及氧化还原态的影响

在培养基中添加不同浓度的硝酸铈和硝酸镧,并检测怀槐细胞悬浮培养过程中异黄酮含量、苯丙氨酸裂解酶(PAL)酶活、还原型谷胱甘肽和氧化型谷胱甘肽含量的变化,以探讨稀土元素对怀槐细胞合成异黄酮的影响和机制.结果表明, 20 mmol L-1硝酸铈和10 mmol L-1硝酸镧较适合诱导怀槐悬浮培养细胞合成异黄酮;细胞培养过程中,硝酸铈和硝酸镧诱导异黄酮合成的变化趋势与PAL酶的活性变化一致;氧化还原态变化分析发现,硝酸铈和硝酸镧处理的细胞中GSH/GSSG比例分别在d 3和d 2达到最大值,是同期对照的1.46和1.47倍,而异黄酮含量则在d 4和d 3达到最大值,分别达到了271.42 μg g-1 (DW)和279.32 μg g-1 (DW).研究结果显示,怀槐细胞悬浮培养中添加稀土元素提高次生代谢产物异黄酮的合成可能与稀土元素诱导细胞氧化还原态的改变有关.图3参16     

水杨酸对小麦高盐毒害的缓解作用

以盐敏感品种中国春、农大８５０２１和抗盐品种冬６８、茶淀红等４个品种小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）为材料,研究盐分胁迫对小麦种子萌发和幼苗生长的影响以及水杨酸对盐害的缓解作用．结果表明,水杨酸能够相对提高盐分胁迫条件下小麦种子萌发的数量、速度和质量;提高幼苗根和叶内游离脯氨酸、可溶性糖等渗透调节物质的含量,增强其渗透调节能力,同时提高幼苗体内超氧化物歧化酶、过氧化物酶等细胞保护酶的活性,抑制膜脂过氧化作用产物丙二醛的积累,降低叶片质膜透性和盐分胁迫对细胞膜的伤害．所有这些变化都有利于缓解盐害,提高小麦抗盐性     

产碱杆菌DN25中降氰酶的分离纯化及生化特性

以一株可降氰的产碱杆菌DN25为酶来源,通过超滤、30 mg/mL硫酸鱼精蛋白沉淀、30%～70%硫酸铵盐析和Phenyl-Toyopearl 650M疏水层析等步骤,获得比活力为44 U/mg的纯化酶制剂.在确定酶浓度、反应时间等氰降解活力测定条件后开展酶学性质研究,试图为将来氰降解代谢机理的深入研究和菌株的基因工程改造提供理论基础.研究结果表明,此纯化酶催化氰化物水解的最适pH值为8.0,最适温度为30℃.该酶在pH 7.0～8.0区域稳定,而在pH>9时会很快失活;在30℃保存10 h,酶活力保持稳定,高于60℃,酶快速失活.加入甘氨酸稳定剂,在60℃下保存20 min酶活仍可保留19.6%.酶促反应动力学符合米氏双曲线方程,测得米氏常数Km为3.11 mmol/L,最大反应速率Vmax为0.23 mmolL-1min-1.     

日本沼虾(Macrobrachium nipponense)N-乙酰-β-D-氨基葡萄糖苷酶初步纯化及部分性质

以日本沼虾内脏为材料,通过硫酸铵沉淀分级分离、DEAE-32离子交换柱层析和SephadexG-100分子筛柱层析纯化,获得比活力为3000Umg-1、纯化倍数为8.88倍的N-乙酰-β-D-氨基葡萄糖苷酶制剂(NAGase).以对-硝基苯-N-乙酰-β-D-氨基葡萄糖为底物,研究酶催化底物水解的反应动力学,结果表明,酶的最适pH为6.0,最适温度为53℃.该酶在pH4.5~9.3区域较稳定,当pH>9.3很快失活;在50℃以下处理30min,酶活力保持稳定,高于50℃,酶稳定性较差,75℃酶完全失活.酶促反应动力学符合米氏双曲线方程,测得米氏常数Km为0.165mmolL-1,最大反应速度Vm为6.55μmolL-1min-1.酶催化pNP-β-D-GlcNAc反应的活化能为63.55kJmol-1.图5表1参16     

米曲霉产氨基酰化酶条件及部分酶学性质研究

通过单因子和多因子摇瓶正交优化试验,确定了米曲霉液态发酵产氨基酰化酶的最佳发酵条件.优化发酵培养基组成(ρ/gL-1):葡萄糖40,蔗糖10,可溶性淀粉20,蛋白胨2.5,马铃薯液1000mL,pH自然.培养基装量50mL/250mL三角瓶,接种量4%.培养温度30℃,转速100r/min,发酵时间42h.每50mL培养物的总酶活由优化前的2627u提高到7338u,是优化前的2.79倍.研究了米曲霉氨基酰化酶的部分酶学性质.该酶催化反应的最适pH为7.0,最适温度为40℃,低浓度的Co2 (5×10-4mol/L)对酶活激活作用显著.图5表2参8     

枯草芽孢杆菌(Bacillus subtilis)过氧化氢酶的分离纯化及性质

对产自枯草芽孢杆菌Bacillus subtilis WSHDZ-01的过氧化氢酶,通过乙醇沉淀、DEAE阴离子交换层析、疏水层析3步纯化,最终获得电泳纯的目标酶(纯化6.8倍).该过氧化氢酶的亚基相对分子质量(M1)为63×103,在405 nm处显示特征吸收峰,推测含有血红素.计算获得酶的表观米氏常数为26.87 mmol L-1.该过氧化氢酶不受低亚硫酸钠的还原作用影响,但被氰化物、叠氮化物和3-氨基-1,2,4-三唑(单功能过氧化氢酶的专一抑制剂)强烈抑制.以邻苯二胺作为电子供体测定酶活时,该酶不显示过氧化物酶活性,因此本文将纯化的过氧化氢酶定性为单功能过氧化氢酶.此外,该酶具有热敏感的特点,且酶活在pH 5～10范围内不受pH影响,此后,活性随着pH的升高而升高,并在pH 11～12处有明显的酶活高峰,在pH 11、25℃放置60 min酶活基本不变,表明该酶在高碱条件下具有很高的活力和一定的稳定性.     

杉木对炭疽病的抗性与苯丙氨酸解氨酶的关系

接种前的杉木（Cunninghamia lanceolata）针叶中，苯丙氨酸解氨酶（PAL）比活力与抗感病性之间不呈规律性相关。不同抗病性的杉木接种炭疽菌（Glomerella cingulata）后，PAL比活力均上升，接种5d后，抗病性强的杉木PAL比活力比抗病性弱的杉木高，接种前以及接种后的各个时间段内，木质素含量都是抗病杉木比感病杉木高。绿原酸的含量与杉木的抗病性之间不呈规律性的相关，从绿原酸对炭疽菌菌丝生长影响的测定结果可知，杉木针叶的绿原酸浓度不足以构成对炭疽菌的影响。可见，接种后针叶的PAL比活力以及接种前、后针叶的木质素含量可作为衡量杉木抗病性的指标。图3表2参14     

茉莉酸类物质(JAs)的生理特性及其在逆境胁迫中的抗性作用

以茉莉酸(JA)和茉莉酸甲酯(MeJA)为代表的茉莉酸类物质(JAs)是广泛存在于高等植物体内的一种新型植物生长调节物质,在调节植物生长发育、光合特性、抗逆反应等起着重要的作用。茉莉酸类物质能抑制植物生长,抑制种子和花粉粒萌发,促进叶片和果实衰老,加速细胞的分裂和膨大,促进气孔关闭,诱导禾本科植物的颖花开放;此外还能调节植物的光合作用和呼吸作用及保护酶活性。目前更多的研究表明,茉莉酸(酯)类物质是与抗性密切相关的植物生长物质,它作为内源信号分子参与植物在机械伤害、病虫害、干旱、盐胁迫、低温等条件下的抗逆反应。在植物受到伤害时,植物体内茉莉酸及其衍生物的含量显著增加,进而诱导一系列与抗逆有关的基因表达,如蛋白酶抑制剂、硫蛋白和苯丙氨酸转氨酶(PAL)等,提高酯氧合酶活性,从而增强植物的抗性。研究还发现,在干旱逆境胁迫条件下,与脱落酸(ABA)的表现相似,茉莉酸类物质大量积累,外施能增强植物的抗旱性。茉莉酸与脱落酸在抑制生长、促进衰老和逆境胁迫的反应上作用极为相似,两者可协同起作用或独立发挥,而茉莉酸与水杨酸(SA)二者之间则大多研究认为存在相互抑制的拮抗作用,但也有一定协同作用。而茉莉酸与各种激素的信号传导途径的相互作用机制仍有待进一步深入研究。     

EDTA-assisted phytoextraction of lead from lead-contaminated soils by Echinochloa crusgalli var. frumentacea

The herbaceous plant Echinochloa crusgalli var. frumentacea is highly resistant to a wide range of heavy metal concentrations. In this study we tested the phytoextraction capacity of E. crusgalli var. frumentacea. Specifically, we compared the effect of EDTA on lead (Pb) accumulation in two groups of plants: those sown in lead contaminated soil and those transplanted to the contaminated soil as seedlings. The result of the time development of the Pb concentrations in the plants in the seedling and seed groups shows that for the seedling group, the effect of adding EDTA to the Pb-contaminated soil was even more pronounced in the shoots than the roots, which showed Pb concentrations 32-fold higher. Compared to the seedling group, the Pb concentrations in the roots of plants in the seed group were approximately 5 times higher in controls and 2 to 10 times higher in the presence of EDTA. Collectively, these results might be considered that EDTA elevates the bioavailability of Pb in soil and this native species is particularly suited to use in Pb phytoextraction.     

化学诱变结合结构类似物法选育高苯丙氨酸解氨酶菌种

ＥＭＳ诱变结合结构类似物法提高含ＰＡＬ出发菌株红酵母Ｒｈｏｄｏｔｏｒｕｌａ ｓｐ．ＣＩＢＡＳＡ１４０１１００ 生物转化能力阳性率为３５．５ ％ ,部分突变株ＰＡＬ转化能力提高１ 倍以上,其中Ｅ１０５,Ｅ２４０ 具有快速转化能力,反应３ ｈＬＰｈｅ累积浓度达到１ ｇＬ左右．φ１０％ 甘油和３０ ｇＬＧｌｕ 对ＰＡＬ转化具有明显的稳定作用．用含φ１０％ 甘油和３０ ｇＬＧｌｕ 的反应液进行生物转化制备,Ｅ１０５ 和Ｅ２４０ 在２４ ｈ 期间ＬＰｈｅ 质量浓度大于２０ ｇＬ,转化率达８０ ％ ．     

污灌与镉胁迫对菠菜几种抗氧化酶活性的影响（Effects of Sewage Irrigation and Cadmium Stresses on the Activities of Several Antioxidant Enzymes of Spinach）

为了探讨污水灌溉与镉胁迫对菠菜抗氧化酶活性的影响,采用盆栽试验的方法,分别用清水和生活污水配制不同浓度(0、1、5、10、50mg·L-1)的Cd2+溶液,对菠菜进行受试处理,每5d灌喷1次,共处理5次.分别采用邻苯二酚比色法、L-苯丙氨酸比色法、水杨酸比色法和高锰酸钾滴定法,对菠菜叶片多酚氧化酶(PPO)、苯丙氨酸解氨酶(PAL)、β-1,3葡聚糖酶(β-1,3glucanase)和过氧化氢酶(CAT)活性进行了测定.结果表明:1)清水处理系列:随着镉处理浓度的增大,与清水对照相比,PPO、PAL活性呈先升高后降低的趋势,CAT活性略有升高,β-1,3葡聚糖酶活性略有降低,各暴露组与清水对照组相比,均呈现显著差异(p<0.05).2)污水处理系列:随着镉处理浓度的增大,与污水对照相比,PPO、PAL活性呈先升高后降低的趋势,CAT和β-1,3葡聚糖酶活性呈逐渐降低趋势;各暴露组与污水对照相比,均呈现显著差异(p<0.05).3)相同Cd2+浓度下,与清水处理相比,各浓度污水处理PAL和PPO活性均明显降低(p<0.05);而β-1,3葡聚糖酶明显升高(p<0.05).以上结果显示:单一镉污染和生活污水与镉复合污染对菠菜不同抗氧化系统酶具有不同影响,这可能与污水中各成分与镉的相互作用有关.     

Impact of cadmium and lead on Catharanthus roseus--a phytoremediation study

The Madagascar Periwinkle, Catharanthus roseus (L.) G. Don (a valued medicinal plant) was exposed to different concentrations ofheavymetals like, CdCl, and PbCl, with a view to observe their bioaccumulation efficiency. Germination was inhibited by both the heavy metals in the seeds previously imbibed in GA, and KNO, for 24 hr. EC50 (the effective concentration which inhibits root length by 50%) was recorded as 180 microM for CdCl2, and 50 microM for PbCl2. Both alpha-amylase and protease activity were reduced substantially on treatment of seeds with increasing concentrations of CdCl2, and PbCl2. Malondialdehyde (MDA) a product of lipoxigenase (LOX) activity also increased due to the treatment of both CdCl, and PbCl2. When two-months-old plants grown in normal soil were transferred to soils containing increasing amounts of these two heavy metals, senescence of lower leaves and extensive chlorosis were noticed after four days of transfer However, plants gradually acclimatized and after 20 days the chlorophyll content was almost comparable to normal. Plants receiving CdCl2 treatment (250 microg g(-1) and less) became acclimatized after two weeks and started normal growth. But PbCl2 of 432 microg g(-1) and less could not affect the plant growth throughout, after a preliminary shock was erased. In case of CdCl2 treatment, a stunted growth with reduced leaf area, reduced biomass and sterility were recorded after six months, while plants show normal growth and flowering in case of PbCl2 treatment. Total alkaloid was also found to be decreased in the roots of CdCl2 treated plants. No change was observed in case of PbCl2. GA3 treatments to the CdCl2 treated plants show internode elongation and increase in leaf area with relatively elongated leaves and thinning of stem diameter AAS analyses of leaves of treated plants exhibited 5-10% accumulation of cadmium, but there was no accumulation of lead at all.     

干旱胁迫对文心兰抗氧化酶活性和渗透调节物质含量的影响

以切花文心兰＂黄金2号＂（Oncidium Gower Ramsey‘Gold 2’）幼苗及成熟苗为材料,用盆栽控制浇水模拟干旱的方法,对不同程度干旱胁迫下其叶片抗氧化酶活性和渗透调节物质含量的变化进行研究。结果表明,文心兰＂黄金2号＂具有一定的耐旱性。随着干旱胁迫程度的加深,文心兰幼苗及成熟苗的相对含水量和相对电导率变化慢、幅度小,超氧化物歧化酶（SOD）和过氧化物酶（POD）活性能保持一定水平,渗透调节物质可溶性糖和脯氨酸含量持续上升,表明文心兰＂黄金2号＂对干旱胁迫具有较强的适应能力。     

Why is latex usually white and only sometimes yellow,orange or red? Simultaneous visual and chemical plant defense

Latex is a toxic and sticky defense substance produced by about 20,000 plant species and secreted following wounding. It defends the wounded plants by both chemical and physical components. A striking fact about latex is that the majority of plants produce it white, although yellow, orange, and red latex can sometimes be found. Theoretically, it is possible that there is no function or importance to latex’s color, but it seems unlikely. It is also possible that there are chemical constraints that dictate the production of white latex, but the various colors indicate that this is not a valid explanation. However, since white latex evolved independently many times in various plant taxa, there should have been a strong selection for latex being white, a possibility that has never been addressed, to my knowledge. I propose that latex is generally white because white is the best solution for visual aposematism under the lighted conditions within and under plant canopies, on the background of typical leaf and bark color, even towards color-blind animals. Yellow, orange, and red latex types should naturally also be considered as aposematic because these are typical colors of toxic and aposematic organisms. In addition, latex-exuding plants simultaneously use a chemically based, non-visual olfactory aposematism because wounding perse also causes the emission of non-latex defensive volatiles and because latex contains various but little-studied defensive volatiles. The volatile defensive aspect of latex exudation may also operate against nocturnal or blind herbivores.     

Development of a cloud condensation nuclei (CCN) counter using a laser and charge-coupled device (CCD) camera

A continuous flow streamwise thermal gradient cloud condensation nuclei (CCN) counter with an aerosol focusing and a laser-charge-coupled device (CCD) camera detector system was developed here. The counting performance of the laser-CCD camera detector system was evaluated by comparing its measured number concentrations with those measured with a condensation particle counter (CPC) using polystyrene latex (PSL) and NaCl particles of varying sizes. The CCD camera parameters (e.g. brightness, gain, gamma, and exposure time) were optimized to detect moving particles in the sensing volume and to provide the best image to count them. The CCN counter worked well in the particle number concentration range of 0.6–8000 #·cm^-3 and the minimum detectable size was found to be 0.5 μm. The supersaturation in the CCN counter with varying temperature difference was determined by using size-selected sodium chloride particles based on K?hler equation. The developed CCN counter was applied to investigate CCN activity of atmospheric ultrafine particles at 0.5% supersaturation. Data showed that CCN activity increased with increasing particle size and that the higher CCN activation for ultrafine particles occurred in the afternoon, suggesting the significant existence of hygroscopic or soluble species in photochemically-produced ultrafine particles.