抗倒胺对大豆幼苗生长的影响（简报）

喷施60～120mg·L－1的抗倒胺（CGR-811）后，大豆幼苗株高明显受抑，基部节间缩短；根系鲜重、干重、根冠比值、根系活力和叶片中叶绿素含量均提高。药剂浓度愈高，其作用愈强，且有良好的壮苗效果。     

氨基酸修饰及荧光光谱分析研究黄花石蒜凝集素生物学活性与构象的关系

本文通过对黄花石蒜凝集素(Lycoris aurea Agglutinin,LAA)进行特殊氨基酸的化学修饰,显示一个LAA分子一共有8个色氨酸分子,其中有3个位于分子表面或近表面,Trp、Tyr和Ser/Thr不是LAA凝集活性所必需的氨基酸,而Asp/Glu的羧基和凝血活性密切相关,对其修饰后导致凝血活性丧失50%.通过荧光淬灭的方法对LAA分子中色氨酸所处微环境进行了研究.结果显示中性淬灭剂丙烯酰胺对LAA分子中色氨酸的淬灭作用最强可以淬灭100%的色氨酸荧光,其次是离子型淬灭剂碘化钾,能淬灭62.9%的色氨酸荧光,而氯化铯对LAA色氨酸的淬灭最弱,几乎不能淬灭LAA的荧光.     

镉和增强紫外线-B辐射复合作用对大豆生长的影响

研究了Cd^2+和增强紫外线-B(UV-B)辐射以及二者复合胁迫(Cd+UV-B)对大豆生长、光合作用、抗氧化酶活性和吲哚乙酸(IAA)氧化酶活性的影响,结果表明,UV-B辐射对大豆生长较CA^2+有更明显的抑制作用,主要是降低了光合作用,生物量减小;抑制节间的分化和伸长,节间减少,株高降低。UV-B辐射对POD、SOD活性有显著诱导作用,而Cd^2+明显颉颃UV-B对POD活性的诱导并抑制IAA氧化酶活性．在复合作用下,植物体内IAA氧化酶和POD活性较UV-B单独作用下显著降低,这两种酶活性降低会引起植物体内IAA含量升高,同时光合作用略有升高,这是株高和生物量较UV-B作用下有所增加的重要原因,复合胁迫还增强了对根伸长生长的抑制作用,根长度较对照显著降低(P<0．05)。IAA氧化酶和POD活性变化以及光合强度变化与大豆株高和生物量变化密切相关,这也是复合胁迫影响大豆生长状况的重要因素。     

半乳糖凝集素-3结合蛋白作为正向调节因子参与IFN-γ介导的抗HBV作用

摘要 目的：研究半乳糖凝集素-3结合蛋白（LGALS3BP）在IFN-?酌介导的抗HBV中的作用。方法：采用qRT-PCR及Western Blot分别检测HepG2细胞瞬转过表达质粒及干扰质粒后LGALS3BP转录水平和翻译水平的变化及检测IFN-γ对HepG2细胞内源性LGALS3BP转录水平和翻译水平的影响；采用CCK-8评价LGALS3BP表达量的变化对HepG2细胞增殖的影响及确定IFN-γ处理HepG2和HepG2.215的最适浓度；采用ELISA检测当LGALSEBP表达量发生变化时，IFN-γ刺激后对细胞外分泌的HBsAg和HBeAg的影响；采用q-PCR和RT-PCR分别检测细胞内HBV核衣壳中DNA和细胞内HBV RNA水平。结果：40 ng/mL的IFN-γ处理HepG2细胞48小时，LGALS3BP mRNA表达量与对照组相比提高2.87倍，但LGALS3BP表达量的变化对细胞增殖无显著作用；当过表达LGALS3BP时，在IFN-γ介导的抗HBV过程中，与对照组相比，HepG2.215细胞分泌的HBsAg、HBeAg以及细胞内HBV DNA和RNA的相对下降幅度分别为46%、67%、59%、49%；与对照组相比，转染pCH9-3091的HepG2细胞分泌的HBsAg、HBeAg以及细胞内HBV DNA和RNA的相对下降幅度分别为67%、53%、60%、29%；而当干扰LGALS3BP时，在IFN-γ介导的抗HBV过程中，与对照组相比，HepG2.215细胞分泌的HBsAg、HBeAg以及细胞内HBV DNA和RNA的相对上升幅度分别为46%、67%、59%、49%；与对照组相比，转染pCH9-3091的HepG2细胞分泌的HBsAg、HBeAg以及细胞内HBV DNA和RNA的相对上升幅度分别为67%、77%、67%、45%。结论：LGALS3BP作为正向调节因子参与IFN-?酌介导的抗HBV过程。     

抗冷冻蛋白基因(afp)转化大豆的研究

目的:通过农杆菌介导法遗传转化大豆。方法:通过热激法将质粒pCAAFP66导入根癌农杆菌菌株EHA105中获得含有抗冷冻蛋白基因(afp)及除草剂抗性筛选标记基因(bar)的农杆菌工程菌株;以大豆品种华春6号和马祖1号种子的下胚轴为外植体,经过农杆菌介导将抗冷冻蛋白基因导入大豆基因组中,在含有除草剂草丁膦(PPT)的培养基中筛选、并经过PCR鉴定获得大豆转化植株。结果:PPT的最佳筛选浓度为1.0mg/L,华春6号和马祖1号的阳性植株数分别为6株和2株,转化效率分别为3.70%和0.94%。结论:不同基因型大豆的转化率存在差异,抗冷冻蛋白基因成功遗传转化进大豆细胞中。     

大豆低聚糖对肠道双歧杆菌和肠杆菌的促生长作用

目的:观察大豆低聚糖和所含糖对肠道菌群中双歧杆菌、肠杆菌体外促生长作用。方法:按1％大豆低聚糖、0．36％水苏糖、0．52％蔗糖、0．11％棉子糖含量配制培养液,分别接种各试验菌株,在0、12、24小时测定各培养液吸光度A值(分光光度计．550nm)和pH值。结果:经24小时培养,加大豆低聚糖的双歧杆菌标准株和临床分离株培养液A值分别为>1．5和1．307,大于肠杆菌标准株和临床分离株的1．1和1．173,而其pH值小于肠杆菌;加水苏糖的双歧杆菌培养液A值为1．47,大于蔗糖的1．4和棉子糖的0．53。结论:大豆低聚糖促双歧杆菌生长作用大于促肠杆菌生长,水苏糖是促双歧杆菌增殖生长的主要成分。     

半夏凝集素基因(pta)导入水稻及其表达的初步研究

半夏凝集素基因是一种有重要价值的抗虫基因。利用RACE—PCR技术克隆出半夏凝集素(pta)基因,并将它构建到载体pCAMBIA1305中形成双元载体(含pta基因和hpt基因)。利用农杆菌介导法将pta基因转入粳稻品种鄂宜105、中花12和籼稻品种E优532成熟胚诱导的愈伤组织中。通过PCR检测,从117株To代再生植株中筛选出36株(其中鄂宜105、中花12、E优532分别为19株、7株、10株)转基因植株。对这些转基因后代植株进行Southern blot分析和RT—PCR分析表明：pta基因已经整合到受体细胞基因组中,并在转录水平上得到了有效表达。通过对转基因后代的遗传分析,成功地从T1代表现为1：2：1盂德尔分离的分离群体中筛选出7个独立转基因水稻纯系(其中包括4个鄂宜105、1个中花12和2个E优532转基因纯系)。对这7个独立转基因水稻E代纯系进行褐飞虱生物抗性鉴定和田间隔离喂养实验,结果显示,这些转基因纯系对褐飞虱的存活率和发育进度均有显著的抑制作用。同时,实验结果还表明2～5mg/L的2,4—D是愈伤组织诱导和生长的必需条件,而且受体的基因型对愈伤组织诱导率和转化频率均有显著影响,在同等条件下,粳稻品种的愈伤组织诱导率和转化频率均高于籼稻种。     

大豆多小叶类型遗传规律初探*

利用合丰25×东农7296系谱法经5个世代选育获得多小叶突变体,并以其作为父本分别与4个小叶正常的栽培大豆配制杂交组合的F1、F2为试验材料,进行多小叶类型的遗传分析。结果表明:小叶数正常的不同大豆亲本与多小叶大豆杂交(3叶×5叶),F1全部植株均表现为5叶,说明5叶性状是受显性核基因控制;不同组合3片叶、3+4片叶、3+5片叶、3+4+5片叶、4+5片叶和5片叶类型组成遗传分离模式存在显著差异,而在3片叶和>3片叶的遗传分离模式相同。杂交F2单株复叶为3片叶和>3片叶的个体分离的比例呈1∶3,符合1对显性单基因的遗传规律。因此,该多小叶突变体牡5796-3的复叶数受1对显性基因控制,该多小叶突变体可作为新种质用于大豆遗传育种及基因克隆和功能研究。     

Light Effect on the Tissue Contents and Distribution of Isoflavones in the Developing Seedling of Soybean

The contents of isoflavones in the developing soybean seedling tissues under different light conditions were examined by high-performance liquid chromatography (HPLC). The results indicated that the total contents of isoflavones in soybean seedling tissues were remarkably different, being the highest in the cotyledons and lower in the leaves and roots. In the cotyledons, the isoflavones like daidzin, genistin and their malonyl conjugates were the major metabolites and were increased markedly with the time of light exposure. In contrast, there was a decrease in the content of isoflavones in the dark. When the dark grown seedlings were tranferred to the light, an increase of the isoflavone content was observed. In the leaves and roots, there was also a marked difference in the contents and types of isoflavones due to various light conditions. Leaves contained mainly the isoflavones such as genistin, malonyl gensistin and flavones rutin, which were increased gradually under light condition, but in the dark, daidzin and malonyl daidzin were the major metabolites and did not change markedly with time. In the roots, no isoflavone was detected in the dark, while five isoflavones were found in the light-grown seedlings, and all root sections were predominated by daidzein and its conjugates. These results provided the evidence that light strongly stimulates the accumulation of isoflavones in soybean seedling tissues.     

大豆多糖对双歧杆菌及人肠道菌群生长的影响

目的研究大豆多糖对双歧杆菌及肠道菌群生长的影响。方法替换Bs培养基中的碳源,分为不加糖、加葡萄糖2％、加大豆多糖2％、加大豆多糖5％、加低聚果糖2％五组,加3种双歧杆菌（长双歧、青春双歧、两歧双歧）菌液1％,测其24h后的活菌数,比较大豆多糖对双歧杆菌生长的影响;替换Bs培养基中的碳源,分为不加糖、加葡萄糖2％、加大豆多糖2％,加低聚果糖2％四组,加人体粪便菌液1％,模拟人体肠道环境厌氧培养24h后,用选择性培养基测其肠杆菌、肠球菌、双歧杆菌、乳酸杆菌的活菌数,观察大豆多糖对人体肠道菌群的影响。结果大豆多糖添加量为5％时对长双歧的促进作用明显优于不加糖组（P〈0．05）;大豆多糖对人体肠道各菌群的生长促进作用与低聚果糖差异无显著性（P〉0．05）。结论大豆多糖对长双歧杆菌的体外促进作用较明显;以粪菌群发酵糖试验表明,大豆多糖对乳杆菌和双歧杆菌均有促进作用,和低聚果糖作用效果相比差异无显著性（P〉0．05）,具有益生元的特性。     

Temporal-spatial co-localisation of tissue transglutaminase (Tgase2) and matrix metalloproteinase-9 (MMP-9) with SBA-positive micro-fibres in the embryonic kidney cortex

Growth of the kidney is a complex process piloted by the collecting duct (CD) ampullae. The dichotomous arborisation and consecutive elongation of this tubular element determines the exact site and time for the induction of nephrons in the overlaying mesenchymal cap condensates. The mechanism by which the CD ampullae find the correct orientation is currently unknown. Recently, we have demonstrated micro-fibres that originate from the basal aspect of the CD ampullae and extend through the mesenchyme to the organ capsule. The micro-fibres are assumed to be involved in the growth and arborisation process of the CD ampulla. Therefore, we have investigated the specific distribution of the micro-fibres during branching morphogenesis. We have also analysed whether the micro-fibres co-localise with extracellular matrix (ECM)-modulating enzymes and whether the co-localisation pattern changes during CD ampulla arborisation. Micro-fibres were detected in all stages of CD ampulla arborisation. Tissue transglutaminase (Tgase2) co-localised with soybean agglutinin (SBA)-positive micro-fibres, whose presence depended upon the degree of CD branching. Matrix metalloproteinase-9 (MMP-9) also co-localised with micro-fibres, but its expression pattern was different from that for Tgase2. Western blotting experiments demonstrated that Tgase2 and MMP-9 co-migrated with SBA-labelled proteins. Thus, the micro-fibres are developmentally modulated by enzymes of the ECM in embryonic kidney cortex. These findings illustrate the importance of micro-fibres in directing CD ampulla growth.     

Cloning and Functional Analysis of the Bifunctional Agglutinin/Trypsin Inhibitor from Helianthus tuberosus L.

In order to find new insect resistance genes, four homologous cDNAs, hta-a, hta-b, hta-c and hta-d with lengths of 775, 718, 784 and 752 bp, respectively （GenBank accession numbers AF477031-AF477034）, were isolated from a tuber cDNA expression library of Helianthus tuberosus L. Sequence analysis revealed that all four cDNAs contain an open reading frame of 444 bp, coding a polypeptide of 147 amino acid residues, and that the sequences of the cDNAs are very similar to those of the mannose-binding agglutinin genes of the jacalin-related family. In hemagglutination reactions and hapten inhibition assays, affinity-purified HTA （Helianthus tuberosus agglutinin） from induced Escherichia coli BL21（DE3） expressing GST-HTA shows hemagglutination ability and a higher carbohydrate-binding ability for mannose than other tested sugars. Trypsin inhibitory activity was detected in the crude extracts of induced E. coli BL21（DE3） expressing HTA, and was further verified by trypsin inhibitory activity staining on native polyacrylamide gel. The mechanism of interaction between HTA and trypsin was studied by molecular modeling. We found that plenty of hydrogen bonds and electrostatic interactions can be formed between the supposed binding sites of HTA-b and the active site of trypsin, and that a stable HTA/trypsin complex can be formed. The results above imply that HTA might be a bifunctional protein with carbohydrate-binding activity and trypsin inhibitory activity. Moreover, Northern blotting analysis demonstrated that hta is predominantly expressed in tubers of H. tuberosus, very weakly expressed in stems, but not expressed at all in other tissues. Southern blotting analysis indicated that hta is encoded by a multi-gene family. The insect resistance traits have been described in another paper.     

Effect of Low Temperature lmbibition on Mito chondrium Respiration and Phosphorylation of PEG Primed Soybean Seed

Cotyledon mitochondrium respiration and oxidative phosphorylation activity of PEG primed and unprimed (control) soybean seeds which have been exposed to low temperature imbibition before germination are studied. The ADP stimulated respiration rates of control mitochondria are evidently higher than state Ⅲ respiration rates of mitochondria from primed seed when L-Mal, α-Kg and Succ are used as substrates respectively. The mitochondria from the unprimed do not possess respiratory control (RC.) On the contrary, mitochondria from the primed, even after seeds being exposed to 2–3 ℃ imbibition for 24 h, phosphorylate normally. The ADP/O and RC values are consistent with those of theoretical expectation. When NADH is used as substrate, unprimed seed mitochondria still possess oxidative phosphorylation activity, while ADP/O and RC values are obviously lower than those of mitochondria from the primed. The emerging sequence of the activity of the diverse phosphorylation sites during germination is also studied. When a different substrate is used, the emerging sequence of the primed is as follows: 1. NADH (12 h), 2.α-Kg (24 h), 3. L-Mal and Succ (48 h). This corresponds to occurrence sequence of ADP stimulated respiration in control mitochondria. The above results show that low temperature imbibition has an irreversible destructive effect on oxidative phosphorylation activity of control mitochondria, and PEG priming has a protective effect on structure and function of the mitochondria under low temperature imbibition stress. The mechanism of soybean imbibitional chilling injury and protective effect of PEG priming are discussed.     

Effect of denervation on the mitotic index of the intestinal epithelium of the rat

Summary The infradiaphragmatic section of vagi nerves at the level of the diaphragm crura causes, on the third day after surgery, a reduction of the mitotic activity in the crypts of the intestinal epithelium of the rat. Moreover there is a drastic reduction in number of the goblet cells that remain concentrated at the lower third of the villi. After the third day after vagotomy there is a tendency to normality.Sympathectomy did not affect significantly the intestinal epithelium of the rat. When both surgeries were performed together, there was no additive effect.     

激光大豆810选育初探

笔者运用激光技术处理大豆萌动种子,从变异后代中育出高产、稳产、优质、多抗、适应怀较强的大豆８１０,缩短了育种年限,是激光技术大豆育种中的重大突破。