蚕豆叶片尿囊素酶的热敏感性

蚕豆叶片尿囊素酶热敏感,在30℃的半衰期是50min。缺底物时预处理10min,半失活温度是41.5℃。甘油能提高其热稳定性,在30％甘油保护下,15℃和25℃之间的活化能是14 644J mol~(-1)(3.5 keal/mol)。     

大豆叶片尿囊素酶的纯化和性质

被纯化的 L—ALNase比活是 819 nkat/mg,酶的纯度提高了738倍,得率为27％。最适pH 7.4,K_m为13.7 mmol/L ALN,对还原剂和巯基试剂敏感,但5 mmol/L的抗坏血酸可提高酶活26％。磷酸盐和KCN有些抑制作用,但Mn~(2+)无活化作用。7.5％ PAGE,ANP定位测定,是 R_f 0.62、0.64、0.68、0.70和 0.725个同功酶的混合物。     

大豆植株中酰脲分布和尿囊素酶活力

大豆品种吉林3号植株不同节位叶片的上部未完全展开叶的酰脲含量和尿囊素酶活性较高。幼茎、叶柄和叶片酰脲含量及尿囊素酶活性高于成年的茎、叶柄和叶片。荚皮酰脲含量显著高于籽粒,但籽粒中的尿囊素酶活性是荚皮中的2～3倍。     

蚕豆植株中酰脲分布和叶片中酰脲水解活性

蚕豆根、茎和叶含有0.31～0.70 μmol酰脲·g~(-1)FW,并受结瘤和生长发育的影响。摘除正在生长的器官可观察到同腋位叶片酞脲含量暂时升高现象。 叶片中酰脲主要是尿囊酸。尿囊素酶和脲酶活性分别为0.30 μmol尿囊酸·g~(-1)FW·h~(-1)和0.19 μmol NH_3·g~(-1)FW·h~(-1)。尿囊酸含量和尿囊素酶活性日变化相似,只是后者峰值比前者出现早。     

赤豆种子萌发中幼苗酰脲含量及尿囊素酶活力变化动态

赤豆种子萌发过程中,幼苗迅速合成酰脲,在酰脲含量达到最大值前,尿囊酸含量高于尿囊素含量,子叶合成酰脲最迅速;酰脲含量达到最大值后,茎带叶的酰脲含量最高。幼苗累积酰脲在品种间相似。种子萌发过程中,幼苗尿囊素酶活力迅速呈线性增加,且与酰脲含量变化趋势一致。幼苗尿囊素酶较耐热。     

二价金属离子在分离大豆种子尿囊素酶中的作用

大豆 (Glycine max L.cv.Keyu 10 )种子和叶片提取液中的尿囊素酶 (ALNase)是热稳定的 ,但在Ca2 、Mg2 或 Mn2 存在的条件下对热有不同的反应。 75℃热处理 5 min,种子提取液中的酶除 Mg2 尚存有 1/ 3酶活性外 ,其余全部丧失 ,56%以上的蛋白质变性 ;叶片提取液中的酶除 Mn2 使活性下降 36%外其余仍然稳定 ,热变性蛋白也不因 Ca2 、Mg2 和 Mn2 的存在有明显增加。在室温下二价金属离子可以清除大豆种子提取液中 50 %以上非酶蛋白 ,但不影响酶活性。二价金属离子这种作用和其浓度无关 ,增加浓度反而有减弱的趋势 ,也不受 Na Cl的影响。两种 ALNase提取液对不同的难溶性钙盐也有不同的反应 ,种子提取液经 5% Ca SO4 处理后非酶蛋白减少近 50 % ,酶活性受影响不大 ;叶片提取液对Ca3 (PO4 ) 2 敏感 ,经处理后酶活性损失 50 %左右 ,蛋白损失近 70 %。 Mn2 对种子提取液中 ALNase有激活作用 ,但这种激活作用随着酶的纯化而消失。乙二胺四乙酸钠 (EDTA)对两种提取液中 ALNase活性没有影响 ,但外加二价金属离子浓度接近 DETA浓度时酶活性很快丢失     

蚕豆豆类胰岛素基因的cDNA克隆及序列分析

为探明豆科植物中豆类胰岛素基因的结构特征与进化关系,在已获得大豆豆类胰岛素基因的基础上,以蚕豆种子胚根mRNA为材料,采用RT-PCR技术,克隆了蚕豆豆类胰岛素基因的cDNA序列,编码的前体多肽包括信号肽、成熟型豆类胰岛素及另一多肽的45个氨基酸残基。DNA序列分析表明,克隆片段与大豆和豌豆的同源性分别为62.5%和58.7%。在氨基酸水平上分别具有44.2%和43.6%的同源性,其中存在着高度保守的半胱氨酸位点,它们在维持豆类胰岛素的空间结构与生理功能方面,可能具有重要的作用。 Abstract:In order to elucidate the relationship between the structural features of leginsulin gene in legume plants and their phylogenetic significance,we have cloned the cDNA sequence of leginsulin gene from radicles of broad bean (Vicia faba) via RT-PCR techniques according to the leginsulin gene sequence we previously obtained from soybean (Glycine max).The cloned cDNA encoded for a precursor protein consisting of the signal peptide,mature leginsulin and an additional 45 amino acids of another polypeptide.A sequence search for homology comparison revealed the cloned leginsulin cDNA fragment shares 62.5% and 58.7% similarity to soybean and pea,respectively.The results also shown that leginsulin cDNA from broad bean presents 44.2% and 43.6% amino acid sequence homology with soybean and pea (Pisum sativum),respectively,and that there exists highly conserved cysteine sites among the leginsulin cDNAs,which may play a crucial role in maintaining the three-dimensional structure and the physiological functions of leginsulin.     

The Composition of the Broad Bean Seed (Vicia faba)

The composition of the seed of the Broad Bean (Vicia faba) hasbeen investigated and the results of alternative analyticalmethods compared. The seeds used in the present work were higherin nitrogen and phosphorus and lower in structural carbohydrate,magnesium and calcium than the seeds used in previous Germananalytical work. Such differences are attributed to differenceof variety and of conditions of maturation of the seed.     

Bee diversity (Hymenoptera: Apoidea) visiting Broad Bean (Vicia faba L.) flowers in Egypt

Different bee flower visitors were observed during the flowering seasons of Broad Bean in Ismailia, eastern Egypt, the Nile Delta and Alexandria, western Egypt. Eight major bee visitors were observed: Andrena ovatula (Kirby, 1802), Andrena sp. (Andrenidae), Chalicodoma siculum (Rossi, 1792) (Megachilidae), Colletes lacunatus Dours, 1872 (Colletidae), Apis mellifera Linnaeus, 1758, Anthophora hispanica (Fabricius, 1787), A. aegyptiaca (Dalla Torre and Friese, 1895) and Xylocopa pubescens (Spinola, 1838) (Apidae). Anthophora hispanica and A. aegyptiaca as well as Chalicodoma siculum had their peak flying period in the middle of the flowering season of the Broad Bean, while Xylocopa pubescens and Andrena sp. had their peak period in the second half of the flowering period.     

蚕豆萎蔫病毒2号分离物侵染对蚕豆叶片光合活性和叶绿体超微结构的影响

研究了受蚕豆萎蔫病毒2号（Broad bean wilt virus 2,BBWV2）中国分离物B935和欧洲分离物PV131侵染的蚕豆（Vicica faba）叶片光合特性、叶绿素荧光诱导动力学参数和叶绿体超微结构变化。感病蚕豆叶绿素含量减少,叶绿素a／b比逐步降低;光合气体交换参数Pn值和Gs值降低,Ci值升高;叶绿素荧光诱导动力学参数Fv/Fm、FV＇/Fm＇、ΦPSII、qP值均有不同程度降低,NPQ值升高;光合器结构遭到不同程度的破坏,B935侵染后叶绿体发育不良,片层结构疏松,PVl31侵染后叶绿体肿胀变圆,片层结构疏松瓦解。与B935相比,PV131侵染对以上各参数的变化有更大影响,且对叶绿体的破坏更为严重。实验结果表明BBWV2不同分离物对光系统II（PSII）的抑制作用与光合器受损程度相关。     

蚕豆叶片发育与衰老过程中超氧物歧化酶活性与丙二醛含量变化

蚕豆植株叶片随茎节自上而下表现出明显的发育与衰老顺序,可作为衰老特征的是叶绿素和蛋白质含量明显下降。蚕豆叶中SOD活性主要定位于12 000× g离心后所得的上清液和叶绿体组分。衰老叶片的SOD总活性和叶绿体组分的相对活性都有所下降,SOD同工酶谱也发生了改变。O_2~ 产生速率随叶龄增大而稍上升;而MDA含量在叶片外观表现枯黄衰老征兆前就急剧上升。可能因为衰老叶片过氧化氢酶活性大幅度下降与SOD之间的不平衡,致使O_2~ 代谢中间产物累积而引起膜的损伤.     

蚕豆根瘤菌接种效应研究

为发掘和利用青海冷凉地区蚕豆优良的根瘤菌种质资源,确定根瘤菌的接种效应。将分离、纯化、分子鉴定的16株蚕豆根瘤菌通过盆栽回接试验的方法进行筛选。结果表明,筛选出6株根瘤菌,它们与青海13号蚕豆共生匹配效果较好,共生固氮能力强,促进蚕豆生长效果明显。     

Axial Resistance to Water Movement in Broad Bean (Vicia faba) Roots

A new potometer method was devised to measure the rates of wateruptake by small portions of an intact root system. An analysisof the relative effects of radial resistance (calculated fromthe potometer data) and axial resistance (calculated from thexylem dimensions) snowed that axial resistance was not likelyto be important. Root water potentials were determined by findingthe osmotic potential of the solution in the potometer whichjust prevented water uptake by the root. Root-tip water potentialswere similar in long and short roots suggesting that any effectof axial resistance was similar in both types of root.     

蚕豆籽粒内的~(14)C同化物的运转和卸出动态

通过向蚕豆叶片饲喂~(14)CO_2,应用液闪和显微放射性自显影技术表明标记同化物经叶脉和果荚韧皮部筛管快速运输至蚕豆种皮。种皮吸收营养、生长,后期逐步降解、供养子叶。种皮内的两类维管束系统同时输送营养并卸出到种皮内侧的质外体空间里。种皮里的反向维管束韧皮部卸出以共质体方式为主。并提供养分供种皮生长,而大部分的同化物由正向完整维管束韧皮部的筛分子一传递细胞进行质外体方式卸出。膨大中的子叶在早期即已成为生理上十分活跃的库。它对标记同化物的摄入随时间进程而急剧上升。