基于离子液体与超声波辅助提取普洱茶茶多酚的工艺优化

本试验利用新型绿色环保溶剂离子液体(1-丁基-3-甲基咪唑溴盐,[BMIM]Br)为提取溶剂,采用超声波辅助提取普洱茶中茶多酚.在以[BMIM]Br浓度、提取时间、提取率及料液比为单因素的试验基础上,进行响应面法优化普洱茶茶多酚的提取工艺.确定最佳提取工艺为:[BMIM]Br浓度0.3 mol/L,料液比1:30,提取时间20 min,超声波功率590 W.在此条件下,普洱茶茶多酚的提取率为8.40％.离子液体与超声波协同作用有效提升了普洱茶茶多酚的提取率,为茶叶中茶多酚的提取工艺提供了新的参考.     

油菜黑胫病菌T-DNA插入诱变因素优化及突变体筛选

油菜黑胫病是由Leptosphaeria biglobosa引起的一种真菌病害。这种病害在我国油菜产区广泛发生，并造成一定的经济损失。为通过获取突变体来研究L. biglobosa的生态适应性机制及致病机制，本文优化了影响农杆菌介导转化（ATMT）油菜黑胫病菌L. biglobosa菌株Lb731的因素，评估转化子质量，并筛选相关突变体。结果明确了农杆菌介导转化菌株Lb731的最佳因素：潮霉素B浓度为50 μg/mL，转化受体（分生孢子）培养时间为15 d (20℃)，浓度为2 × 10^7~8孢子/mL，农杆菌-受体共培养温度为25℃，共培养时间为72 h。在最适条件下的转化效率达到80个转化子/百万分生孢子。T-DNA插入基因组的频率为100%，单拷贝插入频率为72.7%，转化子抗潮霉素性状能稳定遗传。从2136个转化子中获得了11个菌丝生长减缓突变体，7个色素产生缺陷突变体和14个分生孢子产生缺陷突变体，并从这些突变体中鉴定出7个致病力丧失突变体。采用hiTAIL-PCR技术，从3个突变体中获得了T-DNA插入位点侧翼序列。上述结果为深入研究L. biglobosa的生态适应性机制及致病机制提供了材料和线索。     

鸡Ii结合Rab5a和Rab7b分子的分析

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。     

甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物纳米微球作为口蹄疫病毒核酸疫苗递送载体的特性评价

旨在对制备的甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物[poly （D，L-lactide-co-glycolide），PLGA]纳米微球作为口蹄疫病毒（foot-and-mouth disease virus，FMDV）核酸疫苗递送载体进行评价。采用西佛碱反应和元素分析制备具有一定取代度的甘露糖修饰的壳聚糖衍生物（mannose modified chitosan，MCS），然后，经双重乳化挥发法制备得到甘露糖修饰的壳聚糖PLGA纳米微球（MCS-PLGA-NPs）。采用纳米粒径仪检测MCS-PLGA-NPs粒径分布和表面电势（zeta）、扫描电镜考察其形态、琼脂糖凝胶电泳观察其对质粒的吸附和吸附质粒后抵抗核酸酶降解能力、CCK-8法检测MCS-PLGA-NPs的细胞毒性、激光共聚焦观察巨噬细胞对MCS-PLGA-NPs-质粒DNA复合物的摄取、荧光显微镜和Western blot验证MCS-PLGA-NPs加载质粒DNA在细胞中的表达。元素分析结果表明，成功制备了取代度为5%~10%的MCS。纳米粒径测定和扫描电镜结果表明，MCS-PLGA-NPs的zeta为正值、粒径分布均匀且形态规则呈球形。琼脂糖凝胶电泳结果显示，MCS-PLGA-NPs吸附质粒的能力随着其质量的增加而增强并且可以在一定程度上抵抗核酸酶降解质粒DNA。在细胞毒性试验中，不同浓度的MCS-PLGA-NPs与RAW264.7细胞共孵育24 h后，细胞存活率仍在85%以上。在细胞摄取试验中，用激光共聚焦显微镜可以明显观察到质粒DNA结合到纳米微球表面被RAW264.7细胞摄取。荧光显微镜和Western blot试验证明MCS-PLGA-NPs加载质粒DNA可以在细胞中进行表达。综上表明，本研究成功制备了MCS以及具有递送核酸疫苗能力的MCS-PLGA-NPs，为FMDV核酸疫苗的递送研究提供了新的方向和见解，也为该递送载体携带特定抗原靶向抗原递呈细胞表面甘露糖受体以及应用于动物免疫的研究奠定基础。     

安徽省土地资源开发利用成效与保护对策

"十三五"时期,安徽省积极开展土地资源开发利用工作,取得了较大的成效,建设目标基本实现.该文结合"十三五"时期安徽省土地资源开发利用成效,提出了土地资源建设的保护对策,以期为"十四五"期间土地资源的开发利用提供参考.     

高产夏大豆新品种邯豆12的选育及配套栽培技术

邯豆12是邯郸市农业科学院以豫豆25为母本、中黄14(中作92124)为父本进行有性杂交,经过12 a定向选育而成的大豆新品种.2015～2017年参加河北省夏大豆组区域试验、生产试验,邯豆12平均产量为3070.5～3235.3 kg/hm2,较对照增产3.2％～3.9％.该品种成熟时落叶性好,子粒较大,商品性好;中抗花叶病毒病3号株系,感花叶病毒病7号株系.适宜在河北省中南部夏播种植.     

The priority of management factors for reducing the yield gap of summer maize in the north of Huang-Huai-Hai region,China

Understanding yield potential, yield gap and the priority of management factors for reducing the yield gap in current intensive maize production is essential for meeting future food demand with the limited resources. In this study, we conducted field experiments using different planting modes, which were basic productivity(CK), farmer practice(FP), high yield and high efficiency(HH), and super high yield(SH), to estimate the yield gap. Different factorial experiments(fertilizer, planting density, hybrids, and irrigation) were also conducted to evaluate the priority of individual management factors for reducing the yield gap between the different planting modes. We found significant differences between the maize yields of different planting modes. The treatments of CK, FP, HH, and SH achieved 54.26, 58.76, 65.77, and 71.99% of the yield potential, respectively. The yield gaps between three pairs: CK and FP, FP and HH, and HH and SH, were 0.76, 1.23 and 0.85 t ha~(–1), respectively. By further analyzing the priority of management factors for reducing the yield gap between FP and HH, as well as HH and SH, we found that the priorities of the management factors(contribution rates) were plant density(13.29%)fertilizer(11.95%)hybrids(8.19%)irrigation(4%) for FP to HH, and hybrids(8.94%)plant density(4.84%)fertilizer(1.91%) for HH to SH. Therefore, increasing the planting density of FP was the key factor for decreasing the yield gap between FP and HH, while choosing hybrids with density and lodging tolerance was the key factor for decreasing the yield gap between HH and SH.     

Regional distribution of wheat yield and chemical fertilizer requirements in China

Quantification of currently attainable yield and fertilizer requirements can provide detailed information for assessing the food supply capacity and offer data support for agricultural decision-making. Datasets from a total of 5 408 field experiments were collected from 2000 to 2015 across the major wheat production regions in China to analyze the spatial distribution of wheat yield, the soil nutrient supply capacity(represented by relative yield, defined as the ratio of the yield under the omission of one of nitrogen(N), phosphorus(P) and potassium(K) to the yield under the full NPK fertilizer application), and N, P and K fertilizer requirements by combining the kriging interpolation method with the Nutrient Expert Decision Support System for Wheat. The results indicated that the average attainable yield was 6.4 t ha~(-1), with a coefficient of variation(CV) of 24.9% across all sites. The yields in North-central China(NCC) and the northern part of the Middle and Lower reaches of the Yangtze River(MLYR) were generally higher than 7 t ha~(-1), whereas the yields in Southwest China(SWC), Northeast China(NEC), and the eastern part of Northwest China(NWC) were usually less than 6 t ha~(-1). The precentage of area having a relative yield above 0.70, 0.85, and 0.85 for N, P, and K fertilizers accounted for 52.3, 74.7, and 95.9%, respectively. Variation existed in N, P, and K fertilizer requirements, with a CV of 24.8, 23.9, and 29.9%, respectively, across all sites. More fertilizer was needed in NCC and the northern part of the MLYR than in other regions. The average fertilizer requirement was 162, 72, and 57 kg ha~(-1) for N, P_2O_5, and K_2O fertilizers, respectively, across all sites. The incorporation of the spatial variation of attainable yield and fertilizer requirements into wheat production practices would benefit sustainable wheat production and environmental safety.     

不同水肥条件下夏玉米/冬小麦农田生态系统碳平衡研究

农田生态系统碳平衡取决于农作物固定碳量和土壤异养呼吸排放碳量。为揭示水肥用量对农田生态系统碳平衡的综合影响，设置3个灌水水平：高水、中水和低水（W1、W0.85、W0.7夏玉米季分别为90、76.5、63mm，冬小麦季分别为140、119、98mm），4个施氮水平：高氮、中氮、低氮和不施氮（N1、N0.85、N0.7、N0夏玉米季分别为300、255、210、0kg/hm2，冬小麦季分别为210、178.5、147、0kg/hm2），4个施磷水平：高磷、中磷、低磷和不施磷（P1、P0.85、P0.7和P0夏玉米季分别为90、76.5、63、0kg/hm2，冬小麦季分别为150、127.5、105、0kg/hm2）进行了田间试验。结果表明：不同水肥处理下夏玉米/冬小麦农田生态系统表现为碳汇，夏玉米季净生态系统生产力固碳量（CNEP）为6805~7233kg/hm2，冬小麦季CNEP为5842~6434kg/hm2，夏玉米CNEP高于冬小麦。在高、中、低肥水平下，增加灌水量，夏玉米/冬小麦周年净初级生产力固碳量（CNPP）提高2.48%~5.96%，土壤微生物异养呼吸碳释放量（CRm）增加2.15%~15.20%，净生态系统生产力固碳量（CNEP）增加1.16%~6.47%。在高、中、低供水水平下，增加施肥量，夏玉米/冬小麦周年CNPP增加2.95%~3.43%，土壤CRm增加5.23%~18.67%，CNEP增加0.93%~2.79%，CNEP增加比例与供水水平呈负相关。在低水条件下，氮磷肥配施处理夏玉米/冬小麦农田周年CNEP较单施氮、磷肥分别增加4.86%、7.34%，且氮磷肥交互作用显著（P<0.05），水肥供应水平相差15%时对冬小麦农田CNEP有显著的正交互作用。氮磷肥配施、水肥协调供应均有助于促进夏玉米/冬小麦农田生态系统的净碳输入，在节水节肥原则下，夏玉米和冬小麦分别在W0.85N0.85P0.85和W0.7N0.85P0.85水肥供应条件下有利于增加农田CNEP。     

海南狗牙花有效部位提取和体外抗肿瘤活性研究

[目的]初步筛查海南狗牙花抗肿瘤活性部位.[方法]采用MTT比色法测定海南狗牙花4种提取物分别对人非小细胞肺癌细胞(A549)和人乳腺癌细胞(MCF-7)体外生长抑制作用.[结果]海南狗牙花4种提取物对2种肿瘤细胞呈现不同的抑制活性,乙酸乙酯提取物的抗肿瘤活性最强,石油醚提取物次之,正丁醇萃取物较弱而萃取后的水层无抗肿瘤活性.[结论]该研究为深入研究海南狗牙花中具有抗肿瘤活性的单体化合物提供科学依据.