畜禽养殖废水MAP法磷回收试验研究

采用磷酸铵镁法（MAP）回收模拟畜禽养殖废水中的磷,考察了pH值、氮磷比、镁磷以及磷浓度对磷回收率的影响。结果表明,pH是影响MAP结晶沉淀反应产生的重要因素,最佳pH位于9-10;当N/P比超过1∶1,N/P比值变化对P回收率无显著影响,Mg/P对P回收率影响较为明显,适宜的Mg/P比为1∶1左右;磷浓度对磷回收率影响甚小,当磷浓度由5 mg/L增加到150mg/L时,磷回收率均处于99%以上。     

高效聚磷菌株的筛选与鉴定

为了给富营养化水体的净化处理提供参考,对水体中的聚磷菌株进行了筛选和鉴定。结果表明：共筛选到聚磷菌株6株,其中,高效聚磷菌株3株,分别为 E5、E7和 E9,在磷浓度为30 mg／L 培养基中的除磷率分别为96．67％、87．78％和98．90％;通过菌株形态、生理生化特征以及16S rDNA 序列分析,初步鉴定菌株 E5和 E7属于变形菌纲假单孢菌目假单孢菌属的荧光假单孢菌（Pseudomonas fluorescens ）和假单孢菌属某种（Pseudomonas sp）,菌株 E9属于变形菌纲肠杆菌目克雷白氏杆菌属肺炎克雷白氏杆菌肺炎亚种（Klebsiella pneumonia subsp．pneumoniae）。     

‘KRS’芒果发育过程中糖积累与代谢相关酶活性的关系

[目的]探讨芒果果实糖积累和转化的生理机制。[方法]以‘KRS’芒为试材,研究芒果果实生长发育和成熟过程中的淀粉、蔗糖、葡萄糖和果糖含量变化,与淀粉酶、蔗糖代谢相关酶——酸性转化酶(AI)、中性转化酶(NI)、蔗糖合成酶(SS)和蔗糖磷酸合成酶(SPS)活性的相关性。[结果]果实发育前期‘KRS’主要积累淀粉、葡萄糖和果糖,成熟时淀粉酶活性降至最低,淀粉水解快速积累蔗糖。在整个发育过程中AI活性维持最高,完熟时降低,SPS在果实发育中期略有降低,完熟时升至最高,SS和NI一直很低且较稳定。相关性分析表明淀粉含量与酸性转化酶活性呈显著正相关;蔗糖、葡萄糖含量与SPS、SS呈极显著正相关;果糖含量与SS、AI均呈极显著正相关。[结论]芒果成熟时淀粉分解、AI活性降低和SPS、SS活性的增加是引起‘KRS’果实蔗糖积累的主要因子。     

磷酸铵镁堆肥产品养分释放特性及其肥效研究

为了研究磷酸铵镁堆肥产品的氮磷养分释放特性及其肥效,分别采用土柱淋溶方法及盆栽试验研究了磷酸铵镁堆肥(MAPC)产品的氮磷养分释放特性及对油菜的肥效,并与普通化肥(CCH)、普通堆肥(CCO)、磷酸铵镁肥(MAP)3种肥料进行对比。结果表明:在整个淋溶过程中,CCH和MAPC处理淋溶液的总氮及总磷浓度均较其他处理高;MAP肥具有缓释性,且堆肥对其养分释放具有促进作用;CCO处理的养分释放最慢。在最后一次淋洗(即第5次)后,总氮及总磷养分浓度排序均为:CCHMAPCMAPCCO。盆栽试验18 d后油菜快速生长,且与对照(不施肥)处理相比,MAP和MAPC处理能显著提高油菜的株高及生物量。在45 d盆栽试验结束时,CCH、CCO、MAP和MAPC处理的生物量分别是对照处理的14.8、4.8、20.5倍和16.0倍。MAP处理能显著提高油菜的养分含量,TN和TP含量分别比对照提高了2.24 g·kg~(-1)和1.54 g·kg~(-1)。研究表明,4种肥料均可改善油菜的生长发育情况,且MAP和MAPC的肥效优于CCH和CCO,在堆肥中利用鸟粪石结晶原理可固定部分氮磷元素,提高堆肥产品肥效。     

一株柠条内生解磷菌的分离鉴定及实时荧光定量PCR检测

结合形态观察与16S rDNA序列测定对柠条根系内分离筛选得到的解磷细菌C9进行鉴定,Blast比对结果表明C9为泛菌(Pantoea vagans)。在无机磷液体培养基培养条件下,用钼锑抗比色法研究它的解磷能力。结果表明,随着时间的延长,培养液中速效磷含量逐渐增加到4.45mg/L,溶液pH可降至4.2。进一步利用实时荧光定量 PCR 检测柠条根系中、柠条根际土壤与柠条根围土壤中该细菌存在的相对基因拷贝数,结果发现该基因在3种样品中的数量为:柠条根系>柠条根际土壤>柠条根围土壤,表明泛菌解磷细菌能聚集生长在柠条根系内,随着与根系接触距离的增加而呈逐级递减趋势。     

甘蔗B家族蔗糖磷酸合成酶基因SotSPSB的克隆及原核表达

【目的】克隆甘蔗B家族s0心PsB基因并进行原核表达,为进一步研究甘蔗SPS酶学特性及SPS活性调控机制奠定基础。【方法】在进化分析的基础上,通过同源克隆获得甘蔗SofSPSB基因部分序列,再结合RACE技术获得全长cDNA序列。扩增SofSPSB基因ORF并连到原核表达载体pETBlue－2上导入大肠杆菌BL21（DE3）中表达。【结果】通过比对B家族中进化关系很近的玉米（Zeamays）ZmSPS1和水稻（Oryzasativa）OsSPS1基因序列,并在保守区设计一对引物扩增获得甘蔗B家族SPS基因（SolSPSB）2330bp序列。结合5’－RACE和3’－RACE技术获得3481 bp SofSPSB基因全长cDNA序列,该序列包含一个3225bp的开放阅读框（0I江）;起始密码子（ATG）位于转录起始位点后56bp处,终止密码子（TGA）后有一段201bp的非编码序列,并带有真核生物典型的polyA尾巴;编码1074个氨基酸,SofSPSB与Zm—SPS1、OsSPS1的核苷酸序列同源性分别为94．7％和81．3％,氨基酸序列同源性分别为96．0％和83．9％;其理论分子量Mw=118．96kDa,等电点pI=6．30。经原核表达后纯化获得带6xHis标签的融合蛋白。【结论】克隆获得甘蔗B家族Sol-SPSB基因全长cDNA序列,成功构建了SoPSPSB基因原核表达载体,使其在大肠杆菌BL21（DE3）中表达。     

A型肉毒毒素Hc基因的原核表达、纯化及单抗的制备

在大肠埃希茵中高效表达的重组A型肉毒毒素保护性抗原,是以包涵体形式存在.溶解后的包涵体溶液经过处理,用镍柱亲和层析法纯化,得到纯度约为90%的融合蛋白.利用纯化的重组抗原BoNTa免疫Balb/c小鼠,获得分泌抗BoNTa特异性的3株杂交瘤细胞株2A2、4F7和2A8,其单抗鉴定均为IgG1亚型,滴度达到1:105.高纯度和活性单抗的获得为毒素检测试剂盒的研究奠定了基础.     

聚戊烯基磷酸酯的合成机理与化学特征

从银杏叶中分离制备聚戊烯醇混合物(C75~C110),纯度为87.2%,选择POC l3为磷酰化剂和三乙胺为碱性水解剂,经过磷酰化和水解二步反应,聚戊烯基氯磷酸酯在三乙胺碱性溶液中转化成聚戊烯基磷酸酯,产品得率在65%以上。聚戊烯醇与POC l3的摩尔比为1∶5~10,反应温度低于10℃;室温水解20 h,反应产物经柱层析纯化和HPLC制备聚戊烯基磷酸酯,由IR、1H NMR、13C NMR和高分辨质谱(HRMS)鉴定其化学结构为聚戊烯基单磷酸酯。     

DMPP mitigates N2O and NO productions by inhibiting ammonia-oxidizing bacteria in an intensified vegetable field under different temperature and moisture regimes

Vegetable soils with high nitrogen input are major sources of nitrous oxide (N₂O) and nitric oxide (NO), and incorporation of the nitrification inhibitor 3, 4-dimethylpyrazole phosphate (DMPP) into soils has been documented to effectively reduce emissions. However, the efficiency of DMPP in terms of soil N₂O and NO mitigations varies greatly depending on soil temperature and moisture levels. Thus, further evaluations of DMPP efficiency in diverse environments are required to encourage widespread application. A laboratory incubation study (28 d) was established to investigate the interactive effects of DMPP, temperature (15, 25, and 35 ℃), and soil moisture (55% and 80% of water-holding capacity (WHC)) on net nitrification rate, N₂O and NO productions, and gene abundances of nitrifiers and denitrifiers in an intensive vegetable soil. Results showed that incubating soil with 1% DMPP led to partial inhibition of the net nitrification rate and N₂O and NO productions, and the reduction percentage of N₂O production was higher than that of NO production (69.3% vs. 38.2%) regardless of temperature and soil moisture conditions. The increased temperatures promoted the net nitrification rate but decreased soil N₂O and NO productions. Soil moisture influenced NO production more than N₂O production, decreasing with the increased moisture level (80%). The inhibitory effect of DMPP on cumulative N₂O and NO productions decreased with increased temperatures at 55% WHC. Conversely, the inhibitory effect of DMPP on cumulative N₂O production increased with increased temperatures at 80% WHC. Based on the correlation analyses and automatic linear modeling, the mitigation of both N₂O and NO productions from the soil induced by DMPP was attributed to the decreases in ammonia-oxidizing bacteria (AOB) amoA gene abundance and NO^-₂-N concentration. Overall, our study indicated that DMPP reduced both N₂O and NO productions by regulating the associated AOB amoA gene abundance and NO^-₂-N concentration. These findings improve our insights regarding the implications of DMPP for N₂O and NO mitigations in vegetable soils under various climate scenarios.     

Calcium nutrition of the sugarbeet

Abstract

When sugarbeet seedlings are transferred from a complete nutrient solution to one from which Ca has been withheld, the rootlets and tops fail to develop. The same transfer at the eight‐leaf stage causes the rootlets to become stubby and swollen at the tips and blade expansion becomes modified; particularly the upper portions of the blades attaining nearly full development, which pucker and often develop a cupping or hooding effect; a unique symptom characteristic of Ca deficiency. As each new leaf develops, the blade area becomes smaller until only a black tip remains at the apex of the petiole, which is the symptom referred to as tip‐burn for this petiole and the successively . shorter petioles formed as Ca deficiency increases in severity. Strangely, these symptoms also appear during periods of rapid growth when the nutrient solution contains as much as 10 to 28 milliequivalents per liter of Ca or when soils are high in Ca. This implies that Ca absorption and possibly translocation limits the Ca supply at the growing point. Increasing Mg in the nutrient solution decreases Ca uptake and increases Ca deficiency. Potassium deficiency, unexpectedly, induces Ca deficiency apparently by decreasing the translocation of Ca to the growing point.

These phenomena suggest the hypothesis that when ion absorption takes place from the root exchange site that has the affinity for H > Ca > Mg > K > Na, then the H generated internally replaces, and the roots absorb, Na, K and Mg preferentially. Externally, Ca would be adsorbed preferentially from the nutrient solution by the exchange complex, and with the addition of Mg, it would compete for the common adsorption site of Ca and limit Ca absorption internally. Under these conditions potassium‐deficient nutrient solutions would not induce Ca deficiency by decreasing Ca absorption but rather by decreasing Ca translocation. Theoretically, Ba would replace H more readily than Ca on the exchange complex, and therefore, Ba would be adsorbed preferentially and Ca uptake would increase. This effect of Ba was verified experimentally.

Since the translocation of ⁴⁵Ca to the growing point was found to be unrestricted under Ca‐sufficient and Ca‐deficient conditions and since the formation of insoluble Ca compounds such as phosphate or oxalate did not account for the Ca deficiency at the growing point, the cause of the Ca deficiency at the growing point is most likely the higher priority of the storage root for Ca over tops when leaf blades and storage root are both expanding rapidly. However, Ca retransport from older to younger parts of the sugarbeet plant may be restricted by the formation of Ca phosphate under Ca‐deficient conditions and Ca oxalate under Ca‐sufficient conditions.

Calcium deficiency increases net photosynthesis per unit blade area initially, probably because of blade puckering, but not on a per unit chlorophyll basis.