苯丙氨酸解氨酶和绿原酸与小麦抗赤霉病性的关系

对小麦赤霉病不同抗性的苏麦三号(R)、扬麦四号(MR)和南农824(S)三个品种的苯丙氨酸解氨酶(PAL)比活力(u·hr~(-1)·mg~(-1)protein),在拔节期的顶叶中分别测得6.26,12.28和25.12;在孕穗期的麦穗中分别测得140.1,143.3和167.7。滴注接种后8天,苏麦三号、扬麦四号和南农824的病穗中 PAL 比活力与健康穗中的比值分别为0.53,0.67和1.34。用 HPLC 法测定麦穗中绿原酸含量(μg·g~(-1)dry WT),在接种4、8和12天后的苏麦三号穗中分别为32.4,52.4和27.7;扬麦四号穗中分别为60.0,68.0和94.4;南农824穗中分别为123.8,142.2和158.9。绿原酸含量在苏麦三号接种穗中均最低,在南农824接种穗中均最高。禾谷镰刀菌分生孢子在0.05%、0.1%和0.2%的绿原酸溶液中萌发数比对照分别增加214%,614%和970%。上述结果表明,小麦麦穗接种后的 PAL 比活力变化以及绿原酸含量与其抗赤霉病程度呈负相关。     

Purification and characterization of a bifunctional alginate lyase from Pseudoalteromonas sp. SM0524

An alginate lyase-producing bacterial strain, Pseudoalteromonas sp. SM0524, was screened from marine rotten kelp. In an optimized condition, the production of alginate lyase from Pseudoalteromonas sp. SM0524 reached 62.6 U/mL, suggesting that strain SM0524 is a good producer of alginate lyases. The bifunctional alginate lyase aly-SJ02 secreted by strain SM0524 was purified. Aly-SJ02 had an apparent molecular mass of 32 kDa. The optimal temperature and pH of aly-SJ02 toward sodium alginate was 50 °C and 8.5, respectively. The half life period of aly-SJ02 was 41 min at 40 °C and 20 min at 50 °C. Aly-SJ02 was most stable at pH 8.0. N-terminal sequence analysis suggested that aly-SJ02 may be an alginate lyase of polysaccharide lyase family 18. Aly-SJ02 showed activities toward both polyG (α-l-guluronic acid) and polyM (β-D-mannuronic acid), indicating that it is a bifunctional alginate lyase. Aly-SJ02 had lower K(m) toward polyG than toward polyM and sodium alginate. Thin layer chromatography and ESI-MS analyses showed that aly-SJ02 mainly released dimers and trimers from polyM and alginate, and trimers and tetramers from polyG, which suggests that aly-SJ02 may be a good tool to produce dimers and trimers from alginate.     

几种离子对豇豆根瘤腺苷酸琥珀酸裂解酶活性和最适PH值的影响

在含１５％（ｖ／ｖ）甘油和ｐＨ８．０的１００ｍｍｏｌ／ＬＴｒｉｃｉｎｅ缓冲液中，当离子浓度＜１５０ｍｍｏｌ／Ｌ时，Ｋ＋和Ｎａ＋离子是豇豆根瘤腺苷酸琥珀酸裂解酶（ＡＳＡＬ）的激活剂，其最适浓度约为７５ｍｍｏｌ／Ｌ，可分别提高酶活性４０％和３５％。Ｍｇ２＋离子是ＡＳＡＬ的抑制剂，能拮抗Ｋ＋和Ｎａ＋离子的激活效应。ＨＰＯ二４离子可显著降低酶活性，抑制效应与ＨＰＯ二４离子浓度呈正相关。在含１５％（ｖ／ｖ）甘油和ｐＨ６．６～８．０的１００ｍｍｏｌ／Ｌ磷酸盐缓冲液中，ＡＳＡＬ的最适ｐＨ值约为７．４，而在含１５％（ｖ／ｖ）甘油和ｐＨ７．４～８．６的Ｔｒｉｃｉｎｅ缓冲液中，ＡＳＡＬ的最适ｐＨ值约为８．２。     

弱毒菌株尤Ⅱ对小麦条锈病的诱导抗性 Ⅱ.诱导抗性的病生理变化观察

弱小种尤诱导接种后,PAL活性始终大于同期其他处理;先诱导接种尤再挑战接种条中29号处理的POD活性于72h和120h时产生两个高峰;用条中29号小种挑战接种后PPO活性的变化趋势与单独接种尤的处理相同,均高于单独接种条中29号小种和对照处理;挑战接种条中29处理的SOD活性远比同期其他处理的低。     

木薯种质资源中醇腈裂解酶基因12G132600编码区终止突变及频率的研究

木薯是重要的粮食作物,但块根中含有的氰苷及其产生的氢氰酸严重影响木薯的食用品质,增加加工成本。α-醇腈裂解酶（alpha-hydroxy nitrile lyase, HNL）是植物催化醇腈裂解产生氢氰酸的关键酶。重测序数据显示,‘华南8号’木薯品种中HNL编码基因家族的一个成员12G132600在编码区内部发生了终止突变,导致基因编码区变短。本研究对国内外251份木薯种质资源的重测序结果进行分析,共发现60份材料醇腈裂解酶基因12G132600存在相同的终止突变,该60份材料全部是终止突变与非终止突变的杂合基因型,且全部为栽培类型木薯。说明该突变是木薯由野生型驯化为栽培型以后发生的。通过对木薯品种‘华南9号’该基因的克隆测序以及与已发表的木薯品种‘TME3’基因组序列的比对分析,证实了该突变在木薯品种中真实存在。     

锈凹螺褐藻胶裂解酶的分离纯化及性质研究

采用硫酸铵沉淀、离心超滤、DEAE-Sephadex A25、Sephadex G-100和Sephacryl S-300 HR 柱层析等方法从海洋食藻性锈凹螺(Chlorostoma rustica)消化腺中分离纯化出一种褐藻胶裂解酶(Alginate lyase),得到电泳纯酶制品,并对此酶的酶学性质进行了研究.结果表明:此褐藻胶裂解酶的分子量为28 kD,反应的最适温度为40 ℃,具有热不稳定性,最适pH为7.0;酶的反应受多种离子的影响,Mn2 、Co2 和Cd2 对此褐藻胶裂解酶活性具有明显的抑制作用,而Mg2 、Zn2 、Na 和K 则具有促进作用;采用透明圈法和底物法验证了此裂解酶对聚甘露糖醛酸和聚古洛糖醛酸的特异性,实验表明,此酶对聚古罗糖醛酸(Polyguluronate,PG)有特异性.     

SOE法对大豆苯丙氨酸解氨酶(PAL)基因拼接的应用研究

利用SOE(Splicing by Overlap Extension)法对大豆苯丙氨酸解氨酶(PAL)的两个外显子进行拼接,得到完整的PAL基因,表明这种方法是有效可行的.这种技术是作为对cDNA克隆的初步改进与尝试,节约了成本,有一定的应用价值.     

转PAL基因水稻抗稻瘟病性和过氧化物酶活性的研究

 用PAL(苯丙氨酸解氨酶）基因正义和反义转化水稻，获得了70株转基因植株。选择正义转化植株(1s)和反义转化植株(4a)进行稻瘟病菌([i]Magnaporthe grisea[/i])接种，针对病原物侵染，1s的PAL活性上升更快，幅度更大。观察水稻叶片超微结构发现，1s的细胞具有更强的抵抗病原菌入侵的能力，其过氧化物酶活性也比对照和4a要高。     

Separacenes A–D,Novel Polyene Polyols from the Marine Actinomycete,Streptomyces sp

Separacenes A–D (1–4), novel polyene polyols, were isolated from Streptomyces sp. collected from the southern area of Jeju Island, Korea. The chemical structures of 1–4 were established by NMR, mass, UV, and IR spectroscopy as well as the modified Mosher’s method. Separacenes A–B (1–2), which share an identical planar structure but possess different relative configurations, bear tetraene units flanked by two diol moieties, whereas the stereoisomeric separacenes C–D (3–4) possess a triene moiety between two diol substructures. Separacenes A–D each contain a terminal olefinic methylene. Separacene A displayed inhibitory activity against Candida albicans isocitrate lyase and weak cytotoxicity against both the colon carcinoma cell line HCT-116 and the lung cancer cell line A549.