A Novel Gelatinase from Marine Flocculibacter collagenilyticus SM1988: Characterization and Potential Application in Collagen Oligopeptide-Rich Hydrolysate Preparation

Although the S8 family in the MEROPS database contains many peptidases, only a few S8 peptidases have been applied in the preparation of bioactive oligopeptides. Bovine bone collagen is a good source for preparing collagen oligopeptides, but has been so far rarely applied in collagen peptide preparation. Here, we characterized a novel S8 gelatinase, Aa2_1884, from marine bacterium Flocculibacter collagenilyticus SM1988^T, and evaluated its potential application in the preparation of collagen oligopeptides from bovine bone collagen. Aa2_1884 is a multimodular S8 peptidase with a distinct domain architecture from other reported peptidases. The recombinant Aa2_1884 over-expressed in Escherichia coli showed high activity toward gelatin and denatured collagens, but no activity toward natural collagens, indicating that Aa2_1884 is a gelatinase. To evaluate the potential of Aa2_1884 in the preparation of collagen oligopeptides from bovine bone collagen, three enzymatic hydrolysis parameters, hydrolysis temperature, hydrolysis time and enzyme-substrate ratio (E/S), were optimized by single factor experiments, and the optimal hydrolysis conditions were determined to be reaction at 60 ℃ for 3 h with an E/S of 400 U/g. Under these conditions, the hydrolysis efficiency of bovine bone collagen by Aa2_1884 reached 95.3%. The resultant hydrolysate contained 97.8% peptides, in which peptides with a molecular weight lower than 1000 Da and 500 Da accounted for 55.1% and 39.5%, respectively, indicating that the hydrolysate was rich in oligopeptides. These results indicate that Aa2_1884 likely has a promising potential application in the preparation of collagen oligopeptide-rich hydrolysate from bovine bone collagen, which may provide a feasible way for the high-value utilization of bovine bone collagen.     

Design and Synthesis of Novel Phenylahistin Derivatives Based on Co-Crystal Structures as Potent Microtubule Inhibitors for Anti-Cancer Therapy

Phenylahistin is a naturally occurring marine product with a diketopiperazine structure that can bind to the colchicine site of microtubulin as a possible anticancer agent. To develop more potent microtubule inhibitors, novel phenylahistin derivatives were designed and synthesized based on the co-crystal complexes of phenylahistin derivatives and microtubulin. We established a focused library of imidazole-type molecules for the introduction of different groups to the C-ring and A-ring of phenylahistin. Structure–activity relationship studies indicated that appropriate hydrocarbon substituents and unsaturated alkenyl substituents at the 1-position of the imidazole group are important for improving the activity of such compounds. In addition, this study found that propylamine groups could maintain the activity of these compounds, as exemplified by compound 16d (IC₅₀ = 5.38 nM, NCI-H460). Compound 15p (IC₅₀ = 1.03 nM, NCI-H460) with an allyl group exhibited potent cytotoxic activity at the nanomolar level against human lung cancer cell lines. Immunofluorescence assay indicated that compound 15p could efficiently inhibited microtubule polymerization and induced a high expression of caspase-3. 15p also displayed good pharmacokinetic characteristics in vitro. Additionally, the growth of H22 transplanted tumors was significantly inhibited in BALB/c mice when 15p alone was administered at 4 mg/kg, and the tumor inhibition rate was as much as 65%. Importantly, the continuous administration of 15p resulted in a lower toxicity than that of docetaxel (10 mg/kg) and cyclophosphamide (20 mg/kg). Overall, the novel allyl-imidazole-diketopiperazine-type derivatives could be considered safe and effective potential agents for cancer treatment.     

氮肥对大豆不同类型品种结瘤固氮影响的差异性研究

本研究选用生育期和蛋白质含量不同的六个夏大豆品种，在磷钾肥基础上施用三个氮肥水平（０、６、１２ｋｇ氮／亩）。结果表明：施氨抑制结瘤，氮肥越多抑制越严重；抑制作用主要在盛花期前，至鼓粒期有的品种受抑制作用较小；对固氮的抑制在不同品种间存在明显差异，品种豫豆８号表现出较耐氮。酰脲相对丰度与根瘤干重极显著正相关，六叶期、盛花期和鼓粒期两者的相关系数分别为ｒ＿ａ＝０．９５４５、ｒ＿ｂ＝０．８０２５、ｒ＿ｃ＝０．６５３１，Ｐ＜０．０１。酰脲的相对丰度反映了品种的固氮差异和不同氮肥水平下固氮的变化。     

第三次全国农作物种质资源调查与收集行动湖南茶树专项总结报告

2015年10月至2018年12月,项目组参与第三次全国农作物种质资源调查与收集行动,对湖南省茶树种质资源进行普查、调查和征集、收集,共普查、调查和征集、收集茶树种质资源165份,涉及45个农业县(市、区),范围为东经109.176821°~114.005865°、北纬24.943720°~29.819774°、海拔31.33~1469.00m,并对湖南茶树种质资源的保护和合理开发利用提出了建议。     

高寒地区引种籽粒苋研究

在西宁首次引种了５个籽粒苋品种，６年试验结果表明，西宁生态条件下籽粒苋生育期平均１５０ｄ；种子萌发最低温度８￣１０℃，出苗后５０ｄ生长速度加快，在西宁冷凉秋季能够正常灌浆成熟，但播种期提早，生育期则延缓，具有叶面积系数小，干物质生产日积累量小，纱高峰值低以及高峰期短的生长特性。     

免耕整秸秆半覆盖对旱地玉米生长发育及产量的影响

3年试验研究结果表明,旱地玉米采用免耕整秸秆覆盖栽培具有显著的增产效果,在干旱年份尤为明显,3年平均产量比传统耕作玉米增产57.7%,增产幅度23.8%～191.0%.由于秸秆覆盖后土壤温度降低,玉米前期生长发育缓慢,进入拔节期后植株生长发育加快,抽雄前植株长势逐步赶上并超过对照.与传统耕作比较,免耕覆盖玉米具有根系发达、植株高、茎秆粗、叶面积大、干物质积累多、成穗率高、果穗大、穗粒数多、千粒重高等优良性状.     

42%氟啶草酮悬浮剂桶混二甲戊灵对覆膜棉田恶性杂草防除效果及安全性

为了掌握氟啶草酮对覆膜棉田恶性杂草的防除效果及对棉花的安全性,在新疆生产建设兵团南北疆14个团场开展了氟啶草酮与二甲戊灵混用防除棉田龙葵等杂草的田间药效示范试验。结果表明,氟啶草酮与二甲戊灵桶混后于棉花播前土壤封闭处理,对棉田恶性杂草龙葵及灰藜、稗草等一年生阔叶和禾本科杂草均有很好的防除效果,持效期60 d左右,且对棉花出苗和生长安全。     

胡杨果糖-1,6-二磷酸醛羧酶基因PeALD的克隆及耐盐性分析

胡杨（Populus euphratica Oliv）是优良的抗逆树种,有很强的耐盐碱性。前期胡杨转录组研究结果显示盐胁迫可诱导果糖-1,6-二磷酸醛羧酶基因（PeALD）转录上调,提示该基因可能对胡杨耐盐性方面有某种贡献。为分析PeALD基因在胡杨耐盐性中的作用,本研究以胡杨为材料,利用RT-PCR方法克隆了胡杨果糖-1,6-二磷酸醛羧酶基因的全长cDNA,通过基因转化法尝试在烟草中过量表达该基因,并对转基因株系的耐盐性进行初步分析。研究结果显示,克隆的cDNA编码果糖-1,6-二磷酸醛羧酶,ORF为1177bp,是由247个氨基酸编码的疏水性蛋白,理论等电点为6.84,分子量为26.79kD。PCR与RT-PCR检测结果表明,外源的PeALD基因通过转化已经整合到烟草的染色体中,并在4个株系中得到较强的表达。转化株系表型筛选实验表明,在200mmol/L NaCl的MS培养基中培养15d后,野生型烟草植株无明显高生长,叶片全部黄化并萎缩;而转基因烟草高生长基本没有受到抑制,植株生长良好。说明在烟草中过表达PeALD使转化植株的耐盐性显著提高。光合数据结果显示,ALD基因能够降低盐胁迫对烟草叶片净光合速率的影响,可能是PeALD过表达加速了卡尔文循环的运转,提高了CO2的固定速率,进而促进了光合活性,通过光合作用促进碳的积累,利于呼吸作用和氧化磷酸化产生ATP,为维持跨膜质子浓度梯度提供能量,从而有可能促进质膜上的Na＋/H＋逆向转运,利于细胞质膜保持拒盐性。这些结果初步验证了PeALD基因的耐盐性功能,为进一步深入研究该基因在耐盐机制中的作用奠定了良好基础。     

红壤对铝锰离子的吸附特征 Ⅲ.铝锰离子对的交换吸附

对具有水解性质的铝、锰离子对交换吸附研究的结果是 ,两种离子既表现了离子交换的一般化学行为 ,又有明显的非等当量交换和非电性吸附的特征 ;非等当量交换和非电性吸附受制于固相的表面性质、离子对的相对交换能力和离子的加入量 ;对非电性吸附的有关参数k、K、1/n、r和ΔF进行了表述 ;根据铝离子交换锰离子的ΔF ,从热力学论证了红壤对铝离子的吸附是自发和不可逆的并趋向稳态的自然过程     

圈养大熊猫戊型肝炎病毒感染调查

采用ELISA、巢式RT—PCR、免疫组织化学等多种方法对北京动物园保存的大熊猫组织材料和部分现存活个体的粪便、血清等样品进行戊型肝炎病毒抗原调查。调查发现在死亡大熊猫肝脏组织中戊肝病毒抗原阳性检出率为88．9％（8／9）,肾脏的阳性检出率为85．7％（6／7）。现活体动物的粪便、血清抗原检查没有出现阳性结果。结果说明死亡大熊猫曾感染过戊型肝炎病毒,现有个体中没有戊肝病毒感染。