Regulatory role of l-proline in fetal pig growth and intestinal epithelial cell proliferation

l-proline (Pro) is a precursor of ornithine, which is converted into polyamines via ornithine decarboxylase (ODC). Polyamines plays a key role in the proliferation of intestinal epithelial cells. The study investigated the effect of Pro on polyamine metabolism and cell proliferation on porcine enterocytes in vivo and in vitro. Twenty-four Huanjiang mini-pigs were randomly assigned into 1 of 3 groups and fed a basal diet that contained 0.77% alanine (Ala, iso-nitrogenous control), 1% Pro or 1% Pro + 0.0167% α-difluoromethylornithine (DFMO) from d 15 to 70 of gestation. The fetal body weight and number of fetuses per litter were determined, and the small and large intestines were obtained on d 70 ± 1.78 of gestation. The in vitro study was performed in intestinal porcine epithelial (IPEC-J2) cells cultured in Dulbecco''s modified Eagle medium-high glucose (DMEM-H) containing 0 μmol/L Pro, 400 μmol/L Pro, or 400 μmol/L Pro + 10 mmol/L DFMO for 4 d. The results showed that maternal dietary supplementation with 1% Pro increased fetal weight; the protein and DNA concentrations of the fetal small intestine; and mRNA levels for potassium voltage-gated channel, shaker-related subfamily, member 1 (Kv1.1) in the fetal small and large intestines (P < 0.05). Supplementing Pro to either gilts or IPEC-J2 cells increased ODC protein abundances and polyamine concentrations in the fetal intestines and IPEC-J2 cells (P < 0.05). In comparison with the Pro group, the combined administration of Pro and DFMO reduced the expression of ODC protein and spermine concentration in the fetal intestine, as well as the concentrations of putrescine, spermidine and spermine in IPEC-J2 cells (P < 0.05). Meanwhile, the percentage of cells in the S-phase and the mRNA levels of proto-oncogenes c-fos and c-myc were increased in response to Pro supplementation, whereas depletion of cellular polyamines with DFMO increased tumor protein p53 (p53) mRNA levels (P < 0.05). Taken together, dietary supplementation with Pro improved fetal pig growth and intestinal epithelial cell proliferation via enhancing polyamine synthesis.     

低氧胁迫下黄瓜幼苗根系多胺代谢的变化

 利用外源Spd、PAO 抑制剂研究了黄瓜幼苗根系在低氧水培时的生长状况以及内源PAs、H₂O₂ 含量和PAO活性的动态变化。结果表明, 外源PAs对低氧胁迫下黄瓜幼苗根系的生长有促进作用;低氧处理后3种PAs含量都明显上升, 外源Spd和PAO抑制剂增加了根系中的PAs含量, 降低了根系中H₂O₂ 含量, 减缓了低氧伤害; PAs为低氧胁迫的应激产物, PAO为适应酶类, 其活性随PAs含量的变化而发生变化。     

豆梨和杜梨砧木根系中多胺代谢与高温胁迫的关系研究

[目的]探讨豆梨(Pyrus calleryana Dcne)和杜梨(P.betulaefolia Bunge)砧木根系中多胺代谢与其耐高温胁迫的关系,对维持根系正常代谢具有重要意义。[方法]以豆梨和杜梨实生苗为试材,比较了45℃处理下2种砧木叶片落叶率的变化,以及在30和45℃下供试材料根系内源多胺种类、形态和含量变化。[结果]豆梨达到65%落叶率的时间比杜梨早8 d;2种砧木根系中均存在游离态和结合态腐胺、己二胺、亚精胺和精胺4种内源多胺;高温胁迫导致2种供试材料的游离态和结合态腐胺、亚精胺及精胺的含量均升高,其中杜梨根系中游离态腐胺、亚精胺和精胺含量,以及结合态精胺含量显著高于豆梨。[结论]多胺代谢变化与供试材料的耐高温胁迫能力有密切关系。     

玉米弯孢叶斑病菌NADPH氧化酶生物信息学分析和ATMT敲除载体的构建

从JGI数据库玉米弯孢叶斑病菌(Curvularia lunata)基因组中获得2个编码NADPH氧化酶基因ClNox1和ClNox2。采用生物信息学方法对2个基因编码的蛋白质进行性质、结构和功能等方面的预测。结果表明,ClNox1和ClNox2的分子量分别为6.35和6.41 KD,等电点PI为8.87和8.93,不稳定指数为44.50和39.45;ClNox1是亲水性膜蛋白,包含6个明显的跨膜结构域;ClNox2也是膜蛋白,包含8个明显的跨膜结构域。在亚细胞水平上,ClNox1定位于线粒体上,而ClNox2定位于细胞质膜上;在氨基酸序列方面,ClNox1和ClNox2都含有多个苏氨酸、丝氨酸和酪氨酸激酶磷酸化位点;在功能方面,ClNox1和ClNox2都包含NADPH氧化酶特征结构域,并且与Cochliobolus heterostrophus的NADPH氧化酶基因具有较高的同源性。根据ClNox1和ClNox2序列设计特异性引物扩增目的片段,与标记基因NPTII和HphGFP分别连入骨架载体pPZP100,构建完成ClNox1和ClNox2基因的敲除载体pPZP100NPTIIClNox1和pPZP100HphGFPClNox2,为根癌农杆菌介导的遗传转化提供基础材料。     

石竹花芽发生与内源多胺含量的关系

以石竹叶片为外植体,诱导愈伤组织、营养芽、花芽的培养基分别为ＭＳ＋ＺＴ２．５ｍｇ／Ｌ＋２,４－Ｄ０．３ｍｇ／Ｌ;ＭＳ＋ＺＴ１ｍｇ／Ｌ＋ＢＡ２ｍｇ／Ｌ＋ＮＡＡ０．２ｍｇ／Ｌ;ＭＳ＋ＩＢＡ０．５ｍｇ／Ｌ＋ＮＡＡ０．３ｍｇ／Ｌ。在石竹离体叶花芽形成过程中,内源多胺含量呈不同的变化趋势,腐胺、亚精胺含量上升,而二丙胺、精胺含量下降,表明多胺与石竹离体叶花芽分化有明显的内在联系。     

盐胁迫下嫁接伽师甜瓜植株生长与多胺以及多胺氧化酶活性的关系

3种浓度NaCl对嫁接伽师瓜植株生长和产量没有显著抑制作用,而对自根伽师瓜植株的生长和产量产生明显的抑制作用,并随处理浓度的提高抑制作用明显增强。150mmol/L的NaCl胁迫35d嫁接伽师甜瓜植株叶片和根系中的3种形态多胺都有不同程度的下降,其中游离态多胺下降幅度最大;高氯酸不溶性结合态多胺含量变化较小。根系中PAO的活性先上升后下降,而叶片中PAO的活性先下降后上升。游离态多胺中,亚精胺和精胺(Spd+Spm)的含量变化与相应部位PAO的活性变化趋势相反,表明PAO在盐胁迫下可能调节了游离态多胺的含量从而影响高氯酸可溶结合态与高氯酸不溶结合态多胺的含量。     

Genetics and mechanism of resistance to deltamethrin in a field population of Spodoptera litura (Lepidoptera: Noctuidae)

BACKGROUND: Spodoptera litura (F.) causes enormous losses in many economically important crops. The genetics of insecticide resistance has been extensively studied in several insect pests, but there is a lack of information on S. litura. Therefore, the genetics and mechanisms of the resistance of S. litura to deltamethrin were investigated. RESULTS: Bioassays at generation G1 gave resistance ratios of 9, 5, 41, 52 and 49 for deltamethrin, cypermethrin, profenofos, chlorpyrifos and triazofos respectively, when compared with the susceptible Lab-PK strain. Bioassays at G4 with a deltamethrin-selected population (Delta-SEL) showed that selection gave resistance ratios of 63 and 7 for deltamethrin when compared with the Lab-PK and UNSEL strains respectively. Cross-resistance to other insecticides tested was observed in the selected population. A notable feature of the Delta-SEL strain was that resistance to deltamethrin, cypermethrin, profenofos and chlorpyrifos did not decline over the course of five generations. Synergism tests with microsomal oxidase (MO) and esterase-specific inhibitors indicated that the deltamethrin resistance was associated with MO and, possibly, esterase activity. Reciprocal crosses between the Delta-SEL and Lab-PK strains indicated that resistance was autosomal and incompletely dominant. A direct test of monogenic inheritance suggested that resistance to deltamethrin was controlled by more than one locus. CONCLUSION: Stability and dominance of resistance and cross-resistance suggest that insecticides with different modes of action should be recommended to reduce pyrethroid selection pressure.     

渗透胁迫对玉米幼苗叶片不同形态多胺含量的影响

用高压液相色谱法，研究了玉米品种农大108（抗旱性较强）和掖单13（抗旱性较弱）幼苗叶片中游离态(free, f)、酸可溶性共价结合态(AS-conjugated)和酸不溶性共价结合态(AIS-bound) 多胺（polyamine, PA）与渗透胁迫的关系。结果发现，渗透胁迫7 d，农大108的fSpd和fSpm的含量显著(P＜0.05)上升，而掖单13的fPut的升幅显著(P＜0.05)。用S-腺苷蛋氨酸脱羧酶（SAMDC）抑制剂甲基乙二醛-双（鸟嘌呤腙）（MGBG）处理农大108，显著抑制了渗透胁迫诱导的fSpd和fSpm的增加，并且加重了渗透胁迫伤害，用外源Spd处理掖单13，则显著促进了渗透胁迫诱导的fSpd和fSpm的增加，并且减缓了渗透胁迫的伤害。渗透胁迫下，农大108幼苗叶片中AS-conjugated-PA和AIS-bound-PA的上升幅度都显著大于掖单13。用转谷氨酰胺酶（TGase）抑制剂菲咯啉(o-Phen)处理，则抑制了渗透胁迫诱导的AIS-bound-PA含量的上升，并加重了渗透胁迫的伤害。上述结果说明，玉米幼苗叶片中的fSpd、fSpm、AS-conjugated-PA和AIS-bound-PA的升高有利于增强幼苗的抗渗透性。     

水稻胚性悬浮细胞系建立过程中生理生化变化——Ⅱ.氨基酸、多胺及内源激素的变化

以水稻广亲和中粳品系02428为材料,分析了其胚性悬浮细胞系建立过程中氨基酸、多胺及内源激素的变化。结果表明:1.游离氨基酸的总量是FP_(15)>MP_(45)>LP_(90),其中FP_(15)游离精氨酸的含量很高,而在MP_(45)和LP_(90)中含量剧降。蛋白质氨基酸和蛋白质合成能力正好和游离氨基酸的总量变化相反。2.从FP_(15)→MP_(45)→LP_(90),腐胺含量呈上升、精胺含量呈下降趋势。腐胺/(精胺 亚精胺)上升了近20倍。悬浮细胞经脱壁而成游离原生质体后,其总多胺、尸胺和腐胺的含量与悬浮细胞相比均大幅度下降,而精胺和亚精胺则有所升高。3.植物内源激素IAA、2ip的含量是LP_(90)>MP_(45)>FP_(15),ABA、ZRs的含量是FP_(15)>MP_(45)>LP_(90)。上述变化在一定程度上解释了胚性悬浮细胞即后期悬浮细胞适合于进行原生质体培养的生理生化机理。     

稀土及多胺对欧石楠瓶苗生根的效应

本研究以欧石楠继代瓶苗为试材,在生根基质中添加不同浓度的Ce(NO3)3、La(NO3)3和多胺(Spd、Spm、Put),研究其对欧石楠瓶苗生根的影响。结果表明,在欧石楠生根培养基1/2WPM+IBA 0.5 mg/L+NAA 0.2 mg/L中加入0.5～5.0 mg/L Ce(NO3)3和La(NO3)3均可显著提高其生根率、促进根系的伸长生长,而其高浓度对欧石楠瓶苗生根有抑制效应。多胺中的亚精胺(Spd)对其生根具有明显的促进作用,而精胺(Spm)和腐胺(Put)对其生根影响不大。