Crude protein in low‐fish meal diets for juvenile Litopenaeus vannamei can be reduced through a well‐balanced supplementation of essential amino acids

The use of supplemental essential amino acids (EAAs) has been shown to provide an opportunity to minimize excess levels of crude protein (CP) in animal feeds. The present study investigated the effect of reducing the amount of CP in low‐fish meal diets (5%) for juvenile Litopenaeus vannamei. Four sets of diets were prepared containing (% on a fed basis, mean ± SD) 31.24 ± 0.71, 33.70 ± 0.41, 36.90 ± 0.44, and 39.63 ± 0.14% CP, with each protein level containing a total dietary methionine (Met) (Met + Cys) level of 0.56 ± 0.02 (1.07 ± 0.23), 0.71 ± 0.01 (1.22 ± 0.20), 0.88 ± 0.02 (1.38 ± 0.22), or 1.04 ± 0.02% (1.55 ± 0.18%). Shrimp of 1.00 ± 0.08 g were stocked in 84 outdoor tanks of 1 m³ at a rate of 100 shrimp/m² and raised for 75 days. Final survival ranged from 83 to 97% and was unaffected by Met content. Both survival and yield were significantly depressed when shrimp were fed the 31% CP diet. Shrimp grew at a weekly rate of between 0.79 and 0.97 g, achieving a final body weight (BW) in excess of 10.8 g. There was a significant interaction between CP and Met over BW. Shrimp fed 0.56% Met achieved the lowest BW at harvest. Increasing CP beyond 34% did not enhance BW. With a dietary Met content of 0.71%, the highest BW was achieved with 34% CP compared to other levels. There was a significant improvement in Feed Conversion Ratio (FCR) when CP was raised from 31 to 34%. Similarly, dietary Met levels above 0.71% resulted in a significantly better FCR compared to 0.56%. Our study has shown that, if dietary Met (Met + Cys) meets a minimum of 0.71% (1.22%), levels of CP could be reduced from 40 to 34% without adverse effects on shrimp performance.     

生物炭对Bt Cry1Ac蛋白抗紫外能力影响

为了研究生物炭对紫外线的防护作用，以豆壳烧制的生物炭作为载体，研究生物炭对Bt Cry1Ac蛋白的吸附行为以及生物炭对Cry1Ac蛋白的紫外保护作用。使用扫描电子显微镜、透射电子显微镜、X-射线粉末衍射以及傅立叶红外光谱等手段对生物炭的形貌和结构进行表征。结果表明，生物炭是典型的多孔结构材料，表面具有丰富的官能团。Cry1Ac蛋白与生物炭吸附平衡时间为50 min，最合适的吸附浓度比（生物炭：蛋白）为1：100，二者吸附符合准二级动力学模型和Langmuir模型。在UVB紫外照射4 h后，生物炭与Cry1Ac蛋白复合物对棉铃虫的生物活性是单纯蛋白的4.93倍，显示生物炭具有较好的紫外抵抗效果。研究结果初步表明，制备得到的生物炭能够显著提高Cry1Ac蛋白的抗紫外能力，为后续研发耐受紫外线的农药剂型提供新材料。     

灌浆期高温胁迫对不同品种小麦蛋白组分及面团揉混特性的影响

为研究灌浆期高温胁迫对不同品种小麦蛋白组分及面团揉混特性的影响,以济麦22(JM22)和新麦26(XM26)为材料,通过灌浆初期(S1)和灌浆中期(S2)在田间搭棚进行高温胁迫处理,以未进行高温胁迫的大田小麦作对照(CK),收获后对小麦淀粉黏滞谱、蛋白质组分含量和揉混参数等进行分析。结果表明,与各自CK相比,JM22的黏滞谱参数除回复值和糊化温度降低外,其余参数均升高,XM26的黏滞谱参数除峰值时间外均降低。S1和S2使JM22的峰值黏度、低谷黏度、崩解值、最终黏度、峰值时间分别较CK提高2.81%和18.63%、7.71%和19.51%、11.88%和21.15%、1.88%和12.22%、2.45%和4.08%,且S2均大于CK和S1,S1与CK差异不显著;S1和S2使XM26的峰值黏度、低谷黏度、最终黏度、回复值分别较CK降低12.95%和31.21%、1.81%和27.18%、2.50%和22.22%、3.57%和14.39%,其中,S2、S1与CK三者之间的峰值黏度均达显著水平。与CK相比,高温胁迫后JM22的蛋白质含量降低,而XM26升高。两品种各组分蛋白含量均发生改变,S1和S2使JM22谷蛋白含量减少4.52%和6.01%,S1和S2使XM26谷蛋白增加13.66%和17.27%,不可溶蛋白增加28.95%和34.80%,谷醇比也增加。高温处理对两品种面团揉混参数值也有一定影响,S1使JM22和XM26的峰值时间、峰值高度、8 min带宽分别较各自CK显著增加8.50%和42.80%、12.20%和2.80%、26.57%和68.30%,而S2的两品种只有峰值时间同时显著增加,分别增加9.60%和28.50%。本研究结果为优质专用小麦品质提升栽培技术提供了理论基础。     

山葡萄VaCIPK18原核表达与多克隆抗体制备

类钙调磷酸酶B亚基蛋白(CBLs)互作蛋白(CIPKs) 作为类丝氨酸/苏氨酸蛋白激酶,在植物响应非生物胁迫信号转导中起着重要作用。基于前期山葡萄响应低温胁迫转录组测序结果,发现低温胁迫引发的早期伤害及感知阶段的激酶基因中涉及CBL-CIPK 信号通路的VaCIPK18表达显著上调。为进一步研究山葡萄(Vitis amurensis)VaCIPK18激酶参与低温胁迫的功能,采用同源克隆获得了VaCIPK18基因,其开放阅读框为1 320 bp,编码439个氨基酸。基于对VaCIPK18蛋白生物信息学分析,获取胞外结构域中抗原表位丰富的肽段,并将其C端调控结构域(230~439 aa)构建到原核表达载体pET28a-SUMO。将重组表达载体转化至大肠杆菌(E. coli Rosetta)中,经0.8 mmol·L^-1 IPTG、37℃诱导4 h表达出大小为42 kDa的包涵体蛋白。将重组蛋白作为抗原免疫日本大耳白兔,获得anti-VaCIPK18多克隆抗体,经检测具有高效价及特异性。Western Blot结果表明,该抗体可以与葡萄内源CIPK18特异性结合,且在50 kDa位置出现与预期一致的条带。同时,CIPK18在低温胁迫后葡萄叶片中蛋白表达水平与室温下相一致,但两种状态下均存在可能的磷酸化与泛素化修饰现象。本研究结果为进一步探究VaCIPK18的蛋白定位、表达及其功能奠定了基础。     

Application of urease inhibitor improves protein composition and bread‐baking quality of urea fertilized winter wheat

Background: Nitrogen losses is an economic problem for wheat production and a high risk to the environment. Therefore, improved N fertilizer management is a key to increasing the N efficiency and minimizing N losses. To increase N efficiency, enhanced fertilizers such as urea combined with urease inhibitor can be used. Aims: The aim of present study was to evaluate the effects of different N forms on grain storage protein subunits in winter wheat and to examine whether the observed changes correlate with parameters of baking quality. Methods: The investigation was performed over two consecutive years at two locations in Germany. Protein subunits were analyzed by SDS‐PAGE. Results: Protein concentrations were similarly increased after fertilization with ammonium nitrate and urea + urease inhibitor. Analysis of the individual storage protein fractions indicated that both fertilizers specifically enhanced ω‐gliadins and HMW glutenins, but the effect was more pronounced in the ammonium nitrate treatment. Application of urea + urease inhibitor had greater influence on the protein composition and resulted in higher specific baking volume as well as the best fresh keeping ability, in comparison with urea treatment. Conclusion: Considering that the urea + urease inhibitor treatment resulted in almost comparable improvements of NUE and baking quality, with the additional benefit of reduced N losses in combination with easy handling, urea + urease inhibitor can be recommended as a viable alternative to both urea alone and ammonium nitrate treatments. This opens up an opportunity for the reduction of N loss in wheat production when use of urea is preferred.     

江苏省小麦籽粒蛋白质达标弱筋小麦的适生性分析与评价

【目的】弱筋小麦是制作饼干糕点类食品的原料,其烘烤特性很大程度上取决于蛋白质的质和量。小麦籽粒蛋白质含量（GPC,%）不仅由品种的遗传特性决定,还受到气候、土壤、栽培措施等影响。明确江苏省弱筋小麦适宜种植区域以及其地理、气候影响因素,可为江苏弱筋小麦的种植区划提供理论依据。【方法】在2年江苏省小麦品质抽样调查数据的基础上,利用随机森林算法筛选重要性指标,结合单组率Meta分析及其亚组分析,探究地理位置及气象因子对江苏省小麦籽粒蛋白质含量（GPC）达到弱筋小麦标准可能性的影响。【结果】2个年度江苏省小麦GPC平均值为13.92 %,其中2018年、2019年小麦GPC变幅分别为11.06%—18.09%、10.20%—16.50%,平均值分别为14.52%、13.33%,GPC<12.5%的样品分别占比10%、29.71%。从地理分布看,江苏的东南沿湖沿海地区小麦GPC达到弱筋小麦标准的可能性最高,达标可能性最高可达92%,其次是江苏东部沿海地区以及江苏西北部沿河一带。种植地距离一级河流和湖泊或者海岸线的最短距离为20—30 km时,达标可能性相对较高,为23.95%。从气象因子方面看,生育前期特别是出苗期和拔节期,降雨量对江苏弱筋小麦的形成影响较为重要;生育后期尤其是开花期以及灌浆期后期,积温对小麦GPC的影响更重要;且出苗和拔节期的日照时数及开花期的降雨量对江苏弱筋小麦的形成亦很重要,其中,江苏小麦GPC达标弱筋小麦标准的可能性与出苗期的降雨量呈正相关,而与出苗和拔节期的日照时数、拔节期的降雨量以及灌浆后期积温则呈负相关。【结论】江苏弱筋小麦适宜的种植范围受到水系分布与气象因素的共同制约,主要集中在东部沿海和东南沿海沿湖地区。在出苗、拔节期降雨量和开花灌浆期积温适宜的情况下,西北沿河一带的小麦GPC也可达标弱筋小麦标准。品质区划应重点考虑地理位置（水系分布等）和气候分布。     

Effects of ZIP14 on biological behaviors of hepatocellular carcinoma cell line BEL-7404

AIM: To study the expression of zinc transporter ZRT/IRT-like protein 14 (ZIP14) in the hepatocellular carcinoma (HCC) tissues, and to investigate the effects of ZIP14 over-expression on the biological behaviors of HCC cells. METHODS: The expression of ZIP14 at mRNA and protein levels in the HCC tissues and adjacent non-tumor tissues were detected by real-time PCR and immunohistochemical staining, respectively. The lentivirus expression system containing GV365-ZIP14 was constructed, and was used to infect the HCC cell line BEL-7404, which had relatively poor expression of ZIP14. The expression of ZIP14 at mRNA and protein levels in the transfected cells were detected by real-time PCR and Western blot, respectively. Under the conditions of zinc sulfate stimulation at different concentrations, the cell viability, the cell cycle, and the cell migration and invasion abilities were detected by MTT assay, DNA ploid detection, and Transwell assay, respectively. RESULTS: The mRNA expression level and the strong-positive rate of protein expression of ZIP14 in the HCC tissues were significantly lower than those in the adjacent non-tumor liver tissues (P<0.01). The expression of ZIP14 at mRNA and protein levels in the BEL7404 cells was significantly enhanced by infection of GV365-ZIP14 expression lentivirus. Compared with negative control group (transfected with negative control lentivirus), the cell viability, migration and invasion in ZIP14 over-expression group (transfected with GV365-ZIP14 expression lentivirus) were significantly reduced, and the percentage of the cells in G₂/M phase was significantly increased, all of which were more obvious with the elevation of zinc concentration in the culture medium. CONCLUSION: ZIP14 is low expressed in the HCC tissues. The ZIP14 over-expression has inhibitory effects on the viability, migration and invasion of HCC cells, and blocks the cell cycle in G₂/M phase, which might be closely related to the elevation of zinc concentration in cytoplasma of HCC cells due to enchanced zinc transport by ZIP14.     

不同施氮水平下灌浆期高温对水稻贮藏蛋白积累及其合成代谢影响

【目的】 揭示不同施氮水平下灌浆结实期高温对稻米贮藏蛋白积累及其组分的影响,明确不同温氮处理组合下稻米贮藏蛋白合成积累过程与籽粒氮代谢关键酶及相关基因表达间关系。 【方法】 以2个主栽常规晚粳品种（秀水134和秀水09）为材料,利用盆栽土培试验,在水稻穗分化期设低氮（每盆0.5 g尿素）和高氮（每盆2.0 g尿素）2个氮水平,继而通过在水稻灌浆结实期的人工气候箱控温试验,设置高温（日均温度30℃,日最高温和最低温分别为34℃和26℃）和常温（日均温度23℃,日最高温和最低温分别为26℃和20℃）,构成低氮-常温（LN-NT）、低氮-高温（LN-HT）、高氮-常温（HN-NT）、高氮-高温（HN-HT）4个处理组合,并结合水稻籽粒灌浆不同时期取样,探讨氮素穗肥对水稻高温灌浆过程贮藏蛋白积累和主要蛋白组分含量影响及其与籽粒氮代谢关键酶及相关基因表达间关系。 【结果】 氮素穗肥和灌浆期高温均会导致稻米粗蛋白相对含量上升,但在高温处理下单位籽粒中粗蛋白的绝对量却呈降低趋势,以13 kD醇溶蛋白亚基组分在高温处理下的下降幅度最大,从而引起籽粒谷蛋白/醇溶蛋白质比值的上升,其原因主要是由于编码水稻13 kD醇溶蛋白合成基因（Pro13, Pro14和Pro17）在高温处理下的下调表达所致。与之相比,增施氮素穗肥（HN-NT和HN-HT）在引起稻米粗蛋白相对含量提升的同时,单位籽粒中的粗蛋白总量、谷蛋白和醇溶蛋白含量均显著增加,但对贮藏蛋白谷/醇比的影响不明显,且谷蛋白组分中的37 kD酸性亚基和22 kD碱性亚基在不同氮处理水平下的相对比例也基本保持稳定;氮素穗肥可增强灌浆籽粒中的谷氨酰胺合酶（GS）、谷草转氨酶（GOT）和谷丙转氨酶（GPT）活性,但HN-HT处理下的籽粒GS、GOT和GPT活性显著低于HN-NT处理,高温胁迫对水稻灌浆中后期籽粒器官中的氮转运代谢具有抑制作用。此外,在不同氮素水平下,灌浆期高温均可引起稻米整精米率的明显下降和垩白度的显著上升,但HN-HT处理的千粒重、结实率、产量水平、整精米率却高于LN-HT处理,且前者的稻米垩白度显著低于后者,氮素穗肥不足加剧了高温胁迫对千粒重、结实率、整精米率和垩白度等产量和品质性状的负面影响。 【结论】 氮素穗肥对水稻高温灌浆过程贮藏蛋白合成积累起重要调节作用,增施氮素穗肥虽然对水稻籽粒灌浆过程中的谷蛋白和醇溶蛋白质合成具有促进作用,并导致稻米贮藏蛋白相对量与绝对量增加,但对于高温灌浆下13kD醇溶蛋白亚基合成及其组分含量下降具有一定程度的缓解效应,有利于维持贮藏蛋白谷/醇比相对稳定。     

Variation in glomalin in soil profiles and its association with climatic conditions,shelterbelt characteristics,and soil properties in poplar shelterbelts of Northeast China

Glomalin-related soil protein(GRSP)sequesters large amounts of carbon and plays important roles in maintaining terrestrial soil ecosystem functions and ecological restoration;however,little is known about GRSP variation in 1-m soil profiles and its association with stand characteristics,soil properties,and climatic conditions,hindering GRSP-related degraded soil improvement and GRSP evaluation.In this study,we sampled soils from 1-m profiles from poplar(Populus spp.)shelterbelts in Northeast China.GRSP contents were 1.8–2.0 times higher in the upper 40 cm soil layers than at 40–100 cm.GRSP-related soil organic carbon(SOC)sequestration in deeper soil layers was*1.2 times higher than in surface layers.The amounts of GRSP-related nutrients were similar throughout the soil profile.A redundancy analysis showed that in both surface and deeper layers,soil properties(pH,electrical conductivity,water,SOC,and soil nutrients)explained the majority of the GRSP variation(59.5–84.2%);the second-most-important factor in GRSP regulation was climatic conditions(temperature,precipitation,and altitude),while specific shelterbelt characteristics had negligible effects(<5%).Soil depth and climate indirectly affected GRSP features via soil properties,as manifested by structural equation model analysis.Our findings demonstrate that GRSP is important for carbon storage in deep soils,regardless of shelterbelt characteristics.Future glomalin assessments should consider these vertical patterns and possible regulating mechanisms that are related to soil properties and climatic changes.     

大鲵幼体蛋白质的需求量

为评定大鲵幼体对饲料蛋白质的需求量,以鱼粉为主要蛋白源配制6种蛋白质水平(干样基础)的实验饲料:D1(43.7%)、D2(47.1%)、D3(51.3%)、D4(55.7%)、D5(59.9%)和D6(64.4%),饲喂初始体质量为(20.99±0.15)g的大鲵幼体92 d。结果显示,①饲料蛋白质水平对大鲵增重率有显著影响,在D4组达到最大值,较D1组增加了276.4%,且全鲵蛋白质沉积率和肌肉RNA、RNA/DNA值、胃蛋白酶、H^+-K^+-ATPase、胰蛋白酶、脂肪酶和Na^+-K^+-ATPase、肝脏超氧化物歧化酶(SOD)均在D4组达到最佳,而肝脏和肠道丙二醛(MDA)在该组均达到最低;②随饲料蛋白质水平增加,肌肉粗蛋白线性增加,全鲵脂肪线性下降,全鲵水分和粗灰分在各组间差异不显著,全鲵粗蛋白先增加后趋于稳定,在D4组达到最大;③大鲵皮肤胶原蛋白含量在D4组达到最高,较D1组增加了27.83%。研究表明,以增重率、肌肉RNA/DNA值、蛋白质沉积率和皮肤胶原蛋白含量为评价指标,通过二次回归方程分析得到大鲵幼体饲料的最适蛋白质水平为55.9%~58.3%(干样基础),该饲料蛋白质水平能显著提高大鲵幼体胃的泌酸能力、机体消化吸收和抗氧化能力,增加鲵体营养素的沉积,从而促进生长和饲料的转化;而低蛋白质水平饲料显著抑制大鲵的生长。