芹菜水孔蛋白基因AgPIP2;1的克隆及其对非生物胁迫的响应

为了研究芹菜质膜内在蛋白基因的序列特征及其在非生物胁迫条件下的表达特性,探讨其抗逆功能,以芹菜品种六合黄心芹为试验材料,通过RT-PCR技术克隆AgPIP2;1基因,通过生物信息学软件分析其核苷酸和氨基酸序列特征,并采用荧光定量PCR技术检测其在不同组织的表达及其对非生物胁迫的响应。结果表明,AgPIP2;1基因含有1个861 bp的开放阅读框,编码286个氨基酸,其编码的蛋白属于PIP2类蛋白。氨基酸序列分析显示,AgPIP2;1含有高度保守的NPA基序以及高等植物PIP蛋白的保守序列,与其他物种PIP2类蛋白具有较高的同源性,与胡萝卜DcPIP2;1的氨基酸序列同源性达到97.90%。在亲缘关系上与大豆GmPIP2;1、胡萝卜DcPIP2;1和绿豆VrPIP2;1较为接近。实时荧光定量PCR技术分析表明,AgPIP2;1基因在芹菜根、叶柄和叶片中均有表达,其中在叶片中的表达量最高,在根中的表达量最低,呈现比较明显的组织特异性。此外,AgPIP2;1基因受到高温、低温、干旱和盐等非生物胁迫的诱导,说明该基因可能参与芹菜抵御非生物胁迫的过程。本研究为进一步了解AgPIP2;1基因的功能及其在非生物胁迫中的作用奠定了基础。     

大白菜抽薹相关基因BrFLC1的KASP标记开发

为了提高大白菜耐抽薹品种的分子育种效率,本研究针对大白菜抽薹相关基因BrFLC1第6个内含子的第一个碱基(Pi6+1)G-A的SNP变异开发进行高通量检测的竞争性等位基因特异性PCR(KASP)标记。结果表明,开发的KASP标记BrFLC1-KASP1可有效将晚抽薹类型材料Y177-12(G)和早抽薹类型材料Y195-93(A)分为2组。利用BrFLC1-KASP1标记可以对57份大白菜材料的基因型进行有效鉴别,且鉴定结果与酶切扩增多态性标记(CAPS)G-MvaI以及直接测序法的鉴定结果完全一致。综上所述, BrFLC1-KASP1标记在大白菜材料中具有通用性,且具有准确率高、成本低、效率高的特点。本研究结果对大白菜耐抽薹性的分子标记辅助选择具有重要的育种实践价值。     

稳定碳、氮同位素在牛肉加工过程中的分馏效应

为了探究牛肉及副产物中稳定碳、氮同位素在加工过程中的变化规律,确证稳定碳、氮同位素在牛肉加工制品产地溯源中的稳定性和有效性。本试验通过对牛肉进行不同时间的水煮、烤制和油炸3种处理,其中水煮和烤制加工时间分别为5、10、15、20、25和30 min,油炸加工时间分别为1、2、3、4和5 min;采用元素分析仪-稳定同位素比率质谱仪(EA-IRMS)测定脱脂牛肉、粗脂肪及副产物中δ¹³C和δ¹⁵N值。结果表明,脱脂牛肉中δ¹³C值在水煮、烤制和油炸3种加工方式不同加工时间之间均无显著差异,水煮和烤制后粗脂肪中δ¹³C值无显著变化,油炸后的牛肉粗脂肪δ¹³C值主要受植物油的影响,加工方式及加工时间对其无显著影响;水煮脱脂牛肉δ¹⁵N值在加工25、30 min时与对照组牛肉存在显著差异,但平均差值仅为0.3‰~0.9‰。牛肉稳定碳、氮同位素在不同加工过程中分馏效应较小,可有效用于牛肉加工半成品及成品的原产地溯源。     

Intensive organic vegetable production increases soil organic carbon but with a lower carbon conversion efficiency than integrated management

Intensive vegetable production in greenhouses has rapidly expanded in China since the 1990s and increased to 1.3 million ha of farmland by 2016, which is the highest in the world. We conducted an 11‐year greenhouse vegetable production experiment from 2002 to 2013 to observe soil organic carbon (SOC) dynamics under three management systems, i.e., conventional (CON), integrated (ING), and intensive organic (ORG) farming. Soil samples (0–20 and 20–40 cm depth) were collected in 2002 and 2013 and separated into four particle‐size fractions, i.e., coarse sand (> 250 µm), fine sand (250–53 µm), silt (53–2 µm), and clay (< 2 µm). The SOC contents and δ¹³C values of the whole soil and the four particle‐size fractions were analyzed. After 11 years of vegetable farming, ORG and ING significantly increased SOC stocks (0–20 cm) by 4008 ± 36.6 and 2880 ± 365 kg C ha^?1 y^?1, respectively, 8.1‐ and 5.8‐times that of CON (494 ± 42.6 kg C ha^?1 y^?1). The SOC stock increase in ORG at 20–40 cm depth was 245 ± 66.4 kg C ha^?1 y^?1, significantly higher than in ING (66 ± 13.4 kg C ha^?1 y^?1) and CON (109 ± 44.8 kg C ha^?1 y^?1). Analyses of ¹³C revealed a significant increase in newly produced SOC in both soil layers in ORG. However, the carbon conversion efficiency (CE: increased organic carbon in soil divided by organic carbon input) was lower in ORG (14.4%–21.7%) than in ING (18.2%–27.4%). Among the four particle‐sizes in the 0–20 cm layer, the silt fraction exhibited the largest proportion of increase in SOC content (57.8% and 55.4% of the SOC increase in ORG and ING, respectively). A similar trend was detected in the 20–40 cm soil layer. Over all, intensive organic (ORG) vegetable production increases soil organic carbon but with a lower carbon conversion efficiency than integrated (ING) management.     

‘红羽1号’油茶新品种营养成分动态及相关关系分析

为了解‘红羽1号’油茶新品种的营养成分动态及其相关关系，以‘红羽1号’油茶果实近熟期的叶片与果皮、种壳、种仁为材料，测定其还原糖含量、黄酮含量、总酚含量及果皮与叶片中原花青素含量。结果表明，‘红羽1号’油茶中各部位还原糖含量、黄酮含量、总酚含量、原花青素含量具有极显著性差异；在果实近熟期，果实相对成熟，叶片与果皮、种壳、种仁各营养成分动态变化规律明显，但变化幅度不大。叶片原花青素含量与总酚含量间存在显著的线性关系，叶片原花青素含量与黄酮含量和果皮原花青素与总酚含量之间存在显著曲线关系，种壳中总酚含量与黄酮含量极显著正相关。     

Argonaute1敲除前后尖孢镰刀菌转录组分析比较

 Argonaute蛋白(AGO)介导的沉默复合体在RNA干扰(RNAi)中起着至关重要的作用。为了探究AGO1在尖孢镰刀菌RNAi中的作用机制,本文以粘团专化型尖孢镰刀菌生理1号小种FOX-A8野生型和其AGO1缺失突变体(FOX-A8-△Ago1)的菌丝和孢子为材料,分别进行了RNA提取、Illumina HiSeq 2000高通量转录组测序、差异表达基因(DEGs)的显著富集分析;选择菌丝和孢子中的DEGs 进行实时荧光定量PCR(Quantitative real-time PCR, qRT-PCR)验证。GO通路注释结果显示,相对于野生型菌株,AGO1缺失突变体菌丝中的醇脱氢酶(NADP⁺)、孢子中的CAMK / CAMKL / CHK 1蛋白激酶均显著上调;KEGG通路注释结果显示,相对于野生型菌株,AGO1缺失突变体菌丝中与 MAPK信号通路相关的基因、孢子中与PLD信号通路相关的基因均显著下调;另外,相对于野生型菌株,编码AGO2的基因下调,但是下调不显著。qRT-PCR检测DEGs的表达模式与RNA-Seq分析结果一致,证实了RNA-Seq结果的可靠性。     

Trx-1 over-expression attenuates oxidative stress injury in MPP+ -induced PC12 cells by regulating NF-κB signaling pathway

AIM:To investigate the inhibitory effect of thioredoxin 1 (Trx-1) over-expression on oxidative stress injury in 1-methyl-4-phenylpyridinium (MPP⁺)-induced rat pheochromocytoma PC12 cells by regulating NF-κB signaling pathway.METHODS:The PC12 cells were damaged by treatment with MPP⁺ at 1, 3 and 5 mmol/L, and the optimal concentration of 3 mmol/L was selected. The cell viability was measured by MTT assay. The oxidative stress indexes lactate dehydrogenase (LDH) activity, superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the cell culture supernatant were detected, and the protein expression of Trx-1 was determined by Western blot. Lentiviral infection with Ad-Trx-1-GFP sequence was used to establish a model of MPP⁺-treated PC12 cells with Trx-1 over-expression. The effects of Trx-1 over-expression on the cell viability, oxidative stress responses and NF-κB signaling pathway were determined by MTT assay, commercial kits and Western blot. The effects of phorbol 12-myristate 13-acetate (PMA), an activator of NF-κB signaling pathway, on the viability and oxidative stress of PC12 cells were observed. The NF-κB signaling pathway inhibitor pyrrolidine dithiocarbamate (PDTC) was used in MPP⁺-treated PC12 cells with Trx-1 over-expression, and the cell viability and oxidative stress responses were measured. RESULTS:The viability of PC12 cells, SOD activity in the supernatant and protein expression of Trx-1 were decreased, while LDH activity and MDA content in the supernatant were increased significantly by treatment with MPP⁺ at 1, 3 and 5 mmol/L. The effect of MPP⁺ at 3 mmol/L and 5 mmol/L was significantly greater than that at 1 mmol/L (P<0.05), and no significant difference between 3 mmol/L and 5 mmol/L was observed (P>0.05). The inhibitory effect of MPP⁺ on the viability of PC12 cells, and the oxidative stress injury and activation of NF-κB signaling pathway induced by MPP⁺ were significantly attenuated by over-expression of Trx-1. The inhibitory effect of MPP⁺ on the viability of PC12 cells and the oxidative stress injury induced by MPP⁺ were promoted by the activation of NF-κB signaling pathway, while the protective effects of Trx-1 over-expression on the MPP⁺-treated PC12 cells were enhanced by the inhibition of NF-κB signaling pathway. CONCLUSION:Over-expression of Trx-1 protects MPP⁺-treated PC12 cells from oxidative stress injury by regulating NF-κB signaling pathway.     

Effect of FGFR1 expression knock-down on biological characteristics of infantile hemangioma endothelial cells

AIM: To study the effect of fibroblast growth factor receptor 1 (FGFR1) expression knock-down on the viability, apoptosis, invasion and migration of infantile hemangioma endothelial cells (HemECs). METHODS: FGFR1 was down-regulated by FGFR1 small interfering RNA (si-FGFR1) transfection. The viability of the cells was measured by CCK-8 assay. The apoptotic rate was analyzed by flow cytometry and the invasion and migration abilities were determined by Transwell assay. The protein levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and phosphorylated AKT (p-AKT) were examined by Western blot. RESULTS: Transfection of si-FGFR1 into HemECs had significant effects on inhibiting cell viability (P<0.05), promoting apoptosis (P<0.05), and decreasing cell invasion and migration abilities (P<0.05). The results of Western blot showed that knockdown of FGFR1 gene expression in the cells reduced the protein levels of PI3K and p-AKT (P<0.05), and had no significant effect on AKT protein level. CONCLUSION: Knock-down of FGFR1 expression changes the biological characteristics of endothelial cells in infantile hemangiomas by regulating PI3K/AKT signaling pathway.     

基于GF-1卫星数据与面向对象分类的达里诺尔湿地自然保护区土地覆盖信息提取

以达里诺尔湿地自然保护区为研究区，基于国产GF-1遥感影像，采用面向对象和传统目视解译的分类方法对研究区土地覆盖遥感信息进行提取，并对其结果进行对比分析，采取混淆矩阵对面向对象分类结果进行精度验证。结果表明：（1）充分利用了GF-1遥感影像的光谱信息，面向对象分类采取试错法确定最优分割尺度为550，形状和紧致度因子分别为0.6和0.5，各波段权重均为1。（2）面向对象分类总体分类精度达98.22％，KAPPA系数为0.96；（3）面向对象分类方法可快速准确提取类型较为复杂的土地覆盖信息，为内陆湿地精准快速提取研究区土地覆盖分类信息提供参考，以期为湿地遥感业务化监测提供技术规范。     

Neuroprotective effect of novel Rho kinase inhibitor FSD-C10 on a mouse model of Alzheimer disease

AIM: To explore the neuroprotective effect of novel Rho kinase inhibitor FSD-C10 on Alzheimer disease (AD) model of APP/PS1 double transgenic (Tg) mice. METHODS: Male APP/PS1 Tg mice (n=20) at 8 months of age were randomly divided into 2 groups:model group and FSD-C10 treatment group. The mice were treated with normal saline or FSD-C10 (25 mg·kg^-1·d^-1) by intraperitoneal injection, once daily for 2 months. Age-and sex-matched wild-type (WT) mice without treatment were used as the control. The Morris water maze (MWM) test and SMART 3.0 behavioral record system were applied to examine and analyze the spatial cognitive function of the mice. The protein levels and distribution of Aβ, p-tau and synapse-associated proteins such as synaptophysin, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) and postsynaptic density protein 95 (PSD-95) were determined by immunofluorescence staining. The protein levels of phosphorylated amyloid precursor protein at Thr668[p-APP(Thr668)], beta-site APP-cleaving enzyme 1 (BACE1) and synapse-associated proteins in the brain were analyzed by Western blot. RESULTS: Compared with model group, FSD-C10 treatment significantly improved the cognitive function of the APP/PS1 Tg mice, accompanied by reduced Aβ deposition and p-tau level, increased protein level of p-APP (Thr668) in the central nervous system, decreased expression of BACE1, and increased expression of synapse-associated proteins in the brain of the mice (P<0.05). CONCLUSION: FSD-C10 has neuroprotective potential in the APP/PS1 Tg mice. The mechanism may be related to enhancing the non-amyloidogenic APP cleavage pathway, reducing the production of Aβ oligomers, the deposition of senile plaques and the amount of tau protein, up-regulating synapse-associated proteins, and increasing synaptic plasticity.