CRISPR/Cas9技术敲除酿酒酵母gpd2基因对产2,3-丁二醇的影响

旨在利用CRISPR/Cas9基因编辑技术敲除酿酒酵母甘油-3-磷酸脱氢酶基因(gpd2),探究其对2,3-丁二醇产量的影响。根据酿酒酵母(Saccharomyces cerevisiae)W5甘油-3-磷酸脱氢酶基因(gpd2)设计供体片段及gRNA片段,将gRNA片段与可表达Cas9蛋白的敲除载体相连,之后将重组质粒及供体DNA片段转化到S. cerevisiae W5细胞中,根据表型筛选及PCR验证获得gpd2基因缺失菌株。结果表明目的基因gpd2敲除成功,基因缺失菌株与原始菌株经发酵实验相比,甘油产量下降22.01%,乙醇产量提高24.65%,2,3-丁二醇产量下降10.60%。gpd2基因的敲除并没有提高2,3-丁二醇的产量,原因可能是逐渐积累的NADH会优先被细胞内大量的乙醇脱氢酶所氧化,作用于乙醇的产生,而不是优先作用于2,3-丁二醇的合成。本实验构建了适用于酿酒酵母的基因敲除系统,该系统对进一步探究酿酒酵母其他代谢产物与2,3-丁二醇合成之间的关系具有实际的借鉴意义。     

Cf-2/Rcr3pim番茄品系对南方根结线虫的抗性测定

Cf-2/Rcr3~(pim)基因型番茄不仅能够抵御番茄叶霉菌的侵染,而且对马铃薯金线虫的寄生也有一定的抑制效果。为挖掘根结线虫的新抗性资源,本研究采用室内人工接种法测定了Cf-0/Rcr3~(pim)、Cf-2/Rcr3-3和Cf-2/Rcr3~(pim)基因型番茄品系对南方根结线虫的抗感性。抗性评价结果显示,Cf-0/Rcr3~(pim)品系对南方根结线虫表现高感,Cf-2/Rcr3-3品系为中感,而Cf-2/Rcr3~(pim)品系则为感病。与Cf-0/Rcr3~(pim)和Cf-2/Rcr3-3基因型相比,Cf-2/Rcr3~(pim)基因型番茄品系虽然对南方根结线虫侵染的敏感性略低,但是不能阻止线虫在根系上的大量繁殖,不适于根结线虫的防控应用。     

Shikonin inhibits neuronal apoptosis induced by OGD through targeting PI3K/Akt signaling pathway

AIM: To explore the effect of shikonin on rat primary cortical neurons in oxygen-glucose deprivation (OGD)-induced injury model.METHODS: The neurons were pretreated with shikonin at different concentrations (0.02, 0.2, 2 and 20 μmol/L) followed by treatment with OGD. Lactate dehydrogenase (LDH) release assay and fluorescein diacetate/propidium iodide (FDA/PI) double staining were used to detect neuronal viability and apoptosis, and then the optimal concentration of shikonin was determined. LY294002 (PI3K/Akt signaling pathway inhibitor, 1 μmol/L) was added before the addition of shikonin, and the protein level of p-Akt (Ser473) in the neurons was determined by Wes-tern blot. LDH release assay and FDA/PI double staining were also used to detect neuronal viability and apoptosis.RESULTS: A certain concentration (0.2~20 μmol/L) of shikonin increased the viability of impaired neurons (P<0.05) and the protein level of p-Akt (Ser473) in the neurons (P<0.05). The effect of shikonin on neuronal p-Akt (Ser473) levels and the cell death were blocked by LY294002 (P<0.05).CONCLUSION: A certain concentration of shikonin reduces OGD-induced apoptosis of rat primary cortical neurons by activating PI3K/Akt signaling pathway.     

金属3D打印技术的发展现状及制约因素

3D打印技术作为20世纪新兴制造技术,具有高柔性化,生产周期短,材料利用率高等优点,目前在多个领域均有广泛应用。本文综述了目前3D打印技术的发展现状,常用金属3D打印技术的工作原理、特点及发展制约因素等。     

60Co-γ射线对9种庭院月季扦插苗的辐射效应研究

为探讨⁶⁰Co-γ射线对不同庭院月季品种扦插苗的辐射诱变效应,以粉扇、金凤凰、梅朗口红、紫袍玉带、安吉拉、粉色达芬奇、你的眼睛、香槟和九重香9个庭院月季品种的扦插苗为研究对象,⁶⁰Co-γ射线为辐照源,设置不同剂量(0、25、45、60、80 Gy)进行辐射处理,测定了死亡率、萌芽率、变异率和主要的形态指标。结果表明,随着辐射剂量增加,月季扦插苗平均死亡率逐渐增加,从33.61%(0 Gy)上升至79.71%(80 Gy);植株生长受到显著抑制,植株变异率呈先升高后降低的趋势,辐射剂量为45 Gy时,植株变异率达到最高,占总体变异率(4.20‰)的64.70%。此外,不同品种月季对⁶⁰Co-γ射线辐射的敏感性不同,半致死辐射剂量也存在差异,其有效值介于26~55 Gy之间。综合⁶⁰Co-γ射线对月季扦插苗死亡率、生长势和变异率的影响,推荐庭院月季扦插苗的最适辐射剂量为45 Gy,这为月季辐射育种提供了理论基础。     

60Co-γ射线对睡莲生物学效应的影响

为探究辐射对睡莲的生物学效应,对2个睡莲品种弗吉尼亚(Nymphaea Virginia)和奥毛斯特(N.Almost black)的块茎进行不同剂量的⁶⁰Co-γ射线辐射处理。结果表明,2个睡莲品种的存活率均随辐射剂量的增加而下降,半致死剂量分别为24.342和27.671 Gy。5~10 Gy 剂量⁶⁰Co-γ辐射处理使睡莲叶面积和浮叶数显著增加,20~40 Gy剂量⁶⁰Co-γ辐射处理使叶面积和浮叶数显著减少。2个睡莲品种的开花时间均随辐射剂量的增加而延迟,但整个花期长度无显著变化。10~40 Gy剂量⁶⁰Co-γ辐射处理显著降低了睡莲花径,但对睡莲开花率无显著影响。辐射处理导致2个睡莲品种的叶片均出现红棕色斑块、锯齿、皱缩、小孔、卷曲、黄化等变异,且红棕色斑块的面积随着辐射剂量的增加而增大。辐射处理使黑红色花品种奥毛斯特出现普遍的褪色现象,而白色花品种弗吉尼亚的花色未发生变异。此外,经⁶⁰Co-γ射线辐射处理后2个睡莲品种均出现了花型变异的现象。本研究结果为利用辐射诱变技术选育睡莲新品种奠定了基础。     

Biochar addition mitigates nitrogen loss induced by straw incorporation and nitrogen fertilizer application

Biochar has been shown to be potentially beneficial for enhancing yields and soil properties, and diminishing nitrogen (N) losses. However, it remains unclear how biochar regulates soil carbon (C) and N to mitigate N losses induced by straw mixing with N fertilizer in dryland soils. Therefore, we investigated the effects of straw mixing (S₁), S₁ with biochar (SB) and no straw inputs (S₀), and routine urea application rates (N₁) and 70% of routine rates (N_0.7) on yields and N losses, and identify the relationship between N losses and soil C and N compounds. Results showed that N_0.7 and N₁ were suitable for the maize and wheat seasons, respectively, contributing to mitigating N losses without reducing crop yields. Moreover, in the maize season, N_0.7-SB significantly mitigated the straw-induced NH₃-N and N₂O-N emissions by 106% and 81%, respectively. In the wheat season, N₁-SB reduced the straw-induced NH₃-N and N₂O-N emissions by 35% and 66%, respectively. In addition, N_0.7-SB sharply reduced soil inorganic N (SIN) storage in the maize season. Furthermore, the NH₃-N and N₂O-N emission rates were negatively correlated with dissolved organic carbon/SIN content (0–20 cm) (DOC/SIN_0-20). N losses (N₂O-N and NH₃-N emissions and SIN storage) were positively correlated with SIN_0-20, but negatively correlated with soil organic carbon / SIN_0-20 (SOC/ SIN_0-20). This study provides further evidence that biochar with an appropriate N application rate decreased SIN_0-20 and increased DOC/SIN_0-20, thus reducing SIN storage and the straw-induced gaseous N emissions without decreasing crop yields.     

An improved method for extraction of sorghum polymeric protein complexes

A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm.     

秸秆还田与优化施氮改善烤烟根际土壤碳氮转化及氮素吸收

针对华南沙泥田烟区氮肥过量施用导致烟叶品质下降的问题,拟通过秸秆还田与氮肥优化配施调节烤烟各生育期氮吸收、根际土壤碳氮转化及相关酶的活性。试验采用随机区组设计,设水稻秸秆还田与氮肥两因素,其中3个碳（秸秆还田）水平：C0,无秸秆还田,0 kg/hm ²;C1,中量秸秆还田,4500 kg/hm ²;C2,全部秸秆还田,9000 kg/hm ²;2个氮水平：N1,传统施氮,169.50 kg/hm ²;N2,优化施氮,105.00 kg/hm ²。结果表明,与N1相比,N2处理秸秆还田下烤烟前期烟叶和根系氮吸收和累积正常,而后期有所下降;C1、C2处理时,与现蕾期相比,成熟期时根系氮含量均降低4.2 g/kg,叶片氮含量降低3.8、3.0 g/kg;成熟期时,与N1处理相比,N2处理时C1、C2处理的土壤无机氮含量降低30.4%、20.0%,前期2个氮处理间土壤无机氮含量无差异,优化施氮下中量秸秆还田（C1N2）处理烤烟土壤NH₄ ⁺-N含量在整个生育期内较高且呈降低趋势,而硝态氮和土壤无机氮含量在成熟期最低,表明中量秸秆还田下优化施氮能够维持生育前期土壤无机氮含量,且降低成熟期土壤无机氮含量。2个秸秆还田量处理时,团棵期N2处理的土壤转化酶活性是N1处理的1.30、1.13倍;旺长期土壤转化酶活性是N1处理的1.27、1.10倍;与传统施氮相比,秸秆还田下优化施氮处理增加了整个生育期土壤脲酶、磷酸酶、过氧化氢酶活性。在中量秸秆还田（C1）处理下,与N1水平相比,现蕾期和成熟期的N2处理土壤水溶性碳含量提高11.2%和14.1%;与N1相比,N2处理的土壤微生物量氮含量呈降低趋势。综上所述,优化施氮耦合中量秸秆还田（C1N2）能够提高土壤水溶性碳含量,维持土壤氮素合理供应,稳定烤烟生育前期对氮素吸收与累积,减少生育后期烟叶氮素奢侈吸收,与烤烟吸氮规律比较吻合。