小麦TaCYP78A5基因的克隆及生物信息学分析

细胞色素P450(Cytochromes P450)蛋白家族含有硫羟基和血红素结构域,参与多种生物合成,CYP78A家族成员对籽粒大小形成有重要的功能。利用同源克隆的方法,分别从小麦大粒品系‘P271’和小粒材料‘中国春’中扩增到位于2AS、2BS、2DS上的TaCYP78A5-2A、TaCYP78A5-2B和TaCYP78A5-2D基因,编码534、544、549个氨基酸。通过分析发现在‘P271’和‘中国春’克隆到的TaCYP78A5序列一致,说明TaCYP78A5基因在这2个粒型的小麦中是保守的。CYP78A家族在水稻和拟南芥中具有非自主性细胞表达模式,结合对CYP78A家族的生物进化关系研究发现,CYP78A家族的各个基因在单子叶植物中是保守的。‘P271’和‘中国春’籽粒大小的差异,可能是TaCYP78A5基因在‘P271’中的基因表达量高于‘中国春’,引起种子发育过程中内表皮细胞的增殖,从而促进种子表皮增大,使得‘P271’种子大小和质量增加。     

内毒素对大鼠小肠黏膜组织的影响及多价阳离子A的保护效应

旨在揭示大肠埃希菌内毒素(ET)对大鼠小肠黏膜的结构、绒毛长度、上皮内淋巴细胞(IEL)的数量和分布的影响,并探讨多价阳离子A(CA)对上述指标的保护效应。选用72只140g~150g SPF级SD大鼠随机分为3组,即对照组、ET组和CA保护组,经相应处理后分别在3、4、8、12h采集十二指肠、空肠组织作为检测样本,制备病理组织切片,HE染色并利用图像分析系统进行分析。ET组十二指肠和空肠绒毛长度在3、4、8、12h均显著低于对照组和CA保护组(P0.01),ET组十二指肠、空肠IEL数量在3、4、8、12h均显著低于对照组(P0.01);CA保护组十二指肠和空肠IEL数量在4、8、12h均显著高于ET组(P0.01)。结果显示,ET在不同程度上能够破坏小肠黏膜的正常组织结构,降低小肠绒毛长度,减少IEL的数量,从而影响小肠正常的吸收和免疫功能,而CA则能明显降低ET所导致的毒性作用,发挥其保护效应。     

Establishment of Detection Methods for Dexamethasone Sodium Phosphate in Injections by UPLC-PDA

A detection method of dexamethasone sodium phosphate in houttuynia injection,bupleurum injection,vitamin C injection,lincomycin hydrochloride injection,and asragulus polysacharin injection was established by UPLC-PDA.The Waters ACQUITY UPLC^® BEH C18 column(2.1 mm×50 mm,1.7 μm) was used,the mobile phase consisted of 0.75% triethylamine phosphatic liquor/solution (pH 3.0)-methyl alcohol-acetonitrile(50:45:5),the flow rate was 0.3 mL/min,the column temperature had remained 25℃,photodiode array detector wavelength range was 190 to 400 nm,the recorded wavelength was 242 nm,the injection volume was 10 μL.The linear range of dexamethasone sodium phosphatel was 10.0 to 200.0 mg/L (R²=0.9999),the mean recovery and RSD for dexamethasone sodium phosphate in houttuynia injection,bupleurum injection,vitamin C injection,lincomycin hydrochloride injection and asragulus polysacharin injection were 97.54% and 1.14%,101.59% and 0.19%,100.92% and 0.92%,99.82% and 0.73%,100.08% and 0.62%,respectively.This method was accurate,easy and fast,it was suitable for detection of dexamethasone sodium phosphatein houttuynia injection and other four kinds of injections.     

小麦面粉中赭曲霉毒素A的CdSe/ZnS荧光量子点免疫探针检测

构建一种基于荧光量子点免疫探针的小麦面粉中赭曲霉毒素A(OTA)的检测方法。以EDC为活化剂将CdSe/ZnS半导体荧光量子点与OTA单克隆抗体偶联,制备OTA检测探针并评价其检测性能。结果表明:制备的检测探针在3d保存期内具有良好的稳定性。当OTA质量浓度为0.1~0.9μg/L时,检测探针的荧光强度与OTA质量浓度呈线性关系,其相关系数为0.996 3,检出限为0.017 7μg/L。与国标法对照的结果说明,此方法具有可行性。小麦面粉中的加标回收试验结果显示,OTA的回收率为96.0%~102.7%,相对标准偏差不超过4.1%,说明准确性良好。可见,OTA的CdSe/ZnS荧光量子点免疫探针检测方法操作简单、准确度高、结果稳定,有望应用于小麦面粉中OTA的检测。     

Identification of Rosellinia species as producers of cyclodepsipeptide PF1022 A and resurrection of the genus Dematophora as inferred from polythetic taxonomy

Rosellinia (Xylariaceae) is a large, cosmopolitan genus comprising over 130 species that have been defined based mainly on the morphology of their sexual morphs. The genus comprises both lignicolous and saprotrophic species that are frequently isolated as endophytes from healthy host plants, and important plant pathogens. In order to evaluate the utility of molecular phylogeny and secondary metabolite profiling to achieve a better basis for their classification, a set of strains was selected for a multi-locus phylogeny inferred from a combination of the sequences of the internal transcribed spacer region (ITS), the large subunit (LSU) of the nuclear rDNA, beta-tubulin (TUB2) and the second largest subunit of the RNA polymerase II (RPB2). Concurrently, various strains were surveyed for production of secondary metabolites. Metabolite profiling relied on methods with high performance liquid chromatography with diode array and mass spectrometric detection (HPLC-DAD/MS) as well as preparative isolation of the major components after re-fermentation followed by structure elucidation using nuclear magnetic resonance (NMR) spectroscopy and high resolution mass spectrometry (HR-MS). Two new and nine known isopimarane diterpenoids were identified during our mycochemical studies of two selected Dematophora strains and the metabolites were tested for biological activity. In addition, the nematicidal cyclodepsipeptide PF1022 A was purified and identified from a culture of Rosellinia corticium, which is the first time that this endophyte-derived drug precursor has been identified unambiguously from an ascospore-derived isolate of a Rosellinia species. While the results of this first HPLC profiling were largely inconclusive regarding the utility of secondary metabolites as genus-specific chemotaxonomic markers, the phylogeny clearly showed that species featuring a dematophora-like asexual morph were included in a well-defined clade, for which the genus Dematophora is resurrected. Dematophora now comprises all previously known important plant pathogens in the genus such as D. arcuata, D. bunodes, D. necatrix and D. pepo, while Rosellinia s. str. comprises those species that are known to have a geniculosporium-like or nodulisporium-like asexual morph, or where the asexual morph remains unknown. The extensive morphological studies of L.E. Petrini served as a basis to transfer several further species from Rosellinia to Dematophora, based on the morphology of their asexual morphs. However, most species of Rosellinia and allies still need to be recollected in fresh state, cultured, and studied for their morphology and their phylogenetic affinities before the infrageneric relationships can be clarified.     

Insights from in vitro exploration of factors influencing iron,zinc and provitamin A carotenoid bioaccessibility and intestinal absorption from cereals

Developments in genetics, agronomics and processing has positioned staple cereals as important sources of iron, zinc and provitamin A (pVA) carotenoids for nutritionally vulnerable populations. Significant effort has been placed on understanding the bioavailability of these micronutrients from cereal foods, including the exploration of underlying mechanisms by which their bioavailability can be modified. While micronutrient bioavailability is preferably assessed in clinical trials, relevant in vitro digestion and intestinal cell culture models have been applied to study effects of genetic, agronomic, post-harvest and food processing on micronutrient bioavailability. This review (1) critically assesses the application of in vitro models in the exploration of mechanisms associated with iron, zinc and provitamin A carotenoid bioaccessibility and intestinal absorption from cereal foods, and (2) identifies remaining gaps in order to frame future strategies to improve the nutritional impact of cereal foods.     

高粱光敏色素A基因的克隆、蛋白结构与长短日照诱导表达模式

光敏色素是一类红光/远红光受体,对植物光形态建成、外部形态和生理功能等方面起着重要的调节作用。为明晰光敏色素A在调控高粱光形态建成发育过程中的作用,本研究比较分析光温敏感程度不同的高粱品种Btx623、Rio、Tx430和LH645的PhyA基因序列和PHYA蛋白的保守结构域,利用qRT-PCR明确长短日照条件处理下PhyA基因的表达模式。结果表明,Btx623、Rio和Tx430之间PhyA基因序列存在非同义突变,LH645在PhyA基因的第7个外显子上插入了5 bp碱基序列;高粱PhyA蛋白中含有1个PAS-2结构域、1个GAF结构域、1个Phytochrome结构域、2个PAS结构域、1个组氨酸激酶A结构域和1个类似组氨酸激酶的ATP激酶结构;遗传进化树分析表明,单子叶和双子叶植物PhyA蛋白明显聚为2大类,高粱PhyA、谷子PhyA和玉米PhyA1、PhyA2聚为一个亚类,相互间亲缘关系较近,与水稻PhyA同源性相对较远;在短日照(SD)条件下各时期PhyA基因转录水平变化显著,Rio PhyA基因转录水平较Btx623和Tx430变化明显,LH645 PhyA基因转录变化不显著,在长日照(LD)条件下Rio PhyA基因转录水平低于Btx623和Tx430,LH645 PhyA基因转录水平变化不显著。以上结果表明,PhyA受光周期诱导,推测其在高粱光形态建成中起重要调控作用。本研究为进一步解析PhyA基因功能及其在调控高粱光形态建成发育过程中的作用提供理论基础。     

Effect of Psammaplin A on in vitro Development of Bovine Aging Oocytes

To investigate the effect of histone deacetylation inhibitor Psammaplin A (PsA) on the development of bovine aging oocytes in vitro,oocytes were randomly divided into control group,aging group and 50 mmol/L PsA treated aging group (PsA group).Immunofluorescence staining and JC-1 were used to detect the blastocyst rate of bovine oocytes after parthenogenetic activation,the number of cells in blastocysts,apoptosis,reactive oxygen species (ROS),glutathione (GSH) and mitochondrial membrane potential intensity of embryos.The results showed that the blastocyst rate of the aging group was significantly lower than that of PsA and control groups (P<0.05).The blastocyst rate of PsA group was not significantly different from that of control group (P>0.05).The number of cells in the blastocysts of control group and PsA group were significantly higher than that of aging group (P<0.05).The number of cells in the blastocysts of PsA group was not significantly different from that of control group (P>0.05).The apoptosis rate in aging group was significantly higher than that of control and PsA groups (P<0.05),the apoptosis rate of PsA group was significantly higher than that of control group (P<0.05).The GSH level of MⅡ oocytes in aging group was significantly lower than that of control and PsA groups (P<0.05).There was no significant difference in GSH level between control and PsA groups (P>0.05).The ROS level of the embryos in aging group was significantly higher than that of control and PsA groups (P<0.05).The ROS level in PsA group was significantly higher than that of control group (P<0.05).The mitochondrial membrane potential of early embryos of aging group 4-8 cells was significantly lower than that of control and PsA groups (P<0.05).The mitochondrial membrane potential intensity of control group was significantly higher than that of PsA group (P<0.05).In summary,PsA could effectively delay the aging of bovine oocytes and improve the quality of oocytes.     

日粮中添加β-胡萝卜素对牦牛皮下脂肪颜色及组织中维生素A含量的影响

试验旨在探究日粮中添加不同水平β-胡萝卜素（β-C）对牦牛皮下脂肪颜色及组织维生素A含量的影响。试验选取12头2~3岁牦牛为研究对象，随机分为4个组，每组3个重复。各处理组日粮分别在基础日粮中添加0（对照）、720（低剂量）、1 440（中剂量）、1 620 mg/d（高剂量）β-C，饲养90 d后，分别测定牦牛脂肪、肌肉色度、背膘厚度及不同组织中β-C和代谢产物维生素A的含量，结果表明：日粮中添加不同水平β-C，牦牛背部、胸部脂肪及半腱肌的黄度值（b^*）显著高于对照组（P<0.05），亮度值（L^*）和红度值（a^*）没有显著变化（P>0.05）；牦牛背膘厚度在中剂量组达到最大值；对照组牦牛血清中β-C及维生素A含量显著低于各试验组（P<0.05），且维生素A含量随β-C添加量的上升呈上升趋势（P<0.05）；牦牛皮下脂肪、腹腔脂肪、心脏、肝脏、脾脏、肺脏、肾脏及空肠等组织器官中β-C含量在中剂量组均达到最大值，除肝脏中维生素A含量较高外，其他组织中维生素A含量在各处理组间差异不显著（P<0.05）。综上所述，日粮添加β-C影响牦牛皮下脂肪颜色，且在中剂量组牦牛皮下脂肪、心脏、肝脏、脾脏及空肠等组织器官中β-C含量达到最大值，故推荐其最佳摄入量为1 440 mg/d；在日粮中添加β-C影响牦牛肝脏和血清中维生素A含量，对其余组织器官中维生素A含量没有影响。     

质核互作型不育系阜CMS5A大豆的选育及应用

阐述了质核互作型不育系阜CMS5A大豆的选育经过,总结了其特征特性,并分析了其应用成效,以供参考。