分子标记技术在工业大麻性别分化研究中的应用进展

工业大麻性别表达影响着工业大麻种子、纤维和药用成分产量,其性别表达除了受到遗传物质控制外,还受到外界环境条件的影响,这种复杂性使得性别研究一直是工业大麻研究的难点和热点。分子标记技术能在DNA水平上鉴定工业大麻性别,开发与工业大麻性别紧密连锁的分子标记可为大麻性别早期鉴定和特定性别分子辅助选择育种提供理论依据。本文就分子标记在工业大麻性别研究中的应用进展进行了总结,并对应用前景进行了展望。     

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.     

重庆烟区硅酸盐细菌的分离鉴定及生理生化特性

采用土壤稀释平板法从重庆市彭水县的植烟土壤样品中分离出8株具有解钾能力的细菌,通过测定其对含钾矿石中有效钾的释放能力,筛选出两株具有较强的解钾能力的硅酸盐细菌,运用分子生物学方法对这两株硅酸盐细菌的16~23S rDNA序列进行扩增并鉴定。结果表明,一株细菌属于胶质类芽孢杆菌(Paenibacillus mucilaginosus),另一株细菌属于短短芽孢杆菌(Brevibacillus brevis),并且研究了这两株细菌的生理生化特性。     

Variations of SOC and MBC observed in an incubated brown loam soil managed under different tillage systems for 12 years

To assess changes in organic carbon pools, an incubation experiment was conducted under different temperatures and field moisture capacity (FMC) on a brown loam soil from three tillage practices used for 12 years: no‐till (NT), subsoiling (ST) and conventional tillage (CT). Total microbial respiration was measured for incubated soil with and without the input of straw. Results indicated that soil organic carbon (SOC) and microbial biomass carbon (MBC) under ST, NT and CT was higher in soil with straw input than that without, while the microbial quotient (MQ or MBC: SOC) and metabolic quotient (qCO₂) content under CT followed the opposite trend. Lower temperature, lower moisture and with straw input contributed to the increases in SOC concentration, especially under NT and ST systems. The SOC concentrations under ST, with temperatures of 30 and 35°C after incubation at 55% FMC, were greater than those under CT by 28.4% and 30.6%, respectively. The increase in MBC was highest at 35°C for 55%, 65% and 75% FMC; in soil under ST, MBC was greater than that under CT by 199.3%, 50.7% and 23.8%, respectively. At 30°C, the lower qCO₂ was obtained in soil incubated under NT and ST. The highest MQ among three tillage practices was measured under ST at 55% FMC, NT at 65% FMC and CT at 75% FMC with straw input. These data indicate the benefits of enhancing the MQ; the low FMC was beneficial to ST treatment. Under higher temperature and drought stress conditions, the adaptive capacity of ST and NT is better than that of CT.     

转Pi9抗稻瘟病基因水稻株系的比较转录组分析

【目的】在转录水平上解析Pi9基因介导的稻瘟病抗性调控机理,为培育抗病水稻品种提供理论依据。【方法】向水稻品种日本晴(NPB)及其转Pi9抗稻瘟病基因株系(NPB/Pi9)接种稻瘟菌。分别于接种后0 h、12 h、24 h、36 h提取叶组织样品,选取12503个水稻基因定制基因芯片,进行水稻基因转录组分析,并通过qRT-PCR对部分差异表达基因进行验证。【结果】NPB/Pi9在接种后12 h、24 h和36 h的基因表达量分别与其接种0 h表达量比较,共检测到7754个差异表达基因;相应地,感病水稻NPB在以上时间点共检测到7385个差异表达基因;在接种后36 h,NPB/Pi9的差异表达基因数目显著多于NPB。比较NPB/Pi9和NPB相同时间点的基因表达量,共获得4065个差异表达基因,其中接种后36 h的差异表达基因显著多于接种后0 h、12 h或24 h。因此,NPB/Pi9的稻瘟病防御反应更强烈。对NPB/Pi9与NPB相同时间点的差异表达基因进行GO和KEGG分析,细胞外区域、植物对刺激应答、转录调控、氧化还原、离子结合、次生代谢和植物激素相关的GO分类在接种后呈显著富集,苯丙氨酸代谢、类黄酮生物合成和植物激素信号途径的KEGG通路在接种后显著富集。与效应分子触发的免疫反应(ETI)相关的水杨酸信号途径、几丁质酶,以及与病原相关分子模式触发的免疫反应(PTI)相关的胞外区域、对刺激的应答、木质素合成等,均在抗感水稻之间差异表达。而且PTI/ETI共有的WRKY转录因子、MAPK激酶、茉莉酸和乙烯信号途径等发生差异表达。综上所述,NPB/Pi9和NPB的差异表达模式与ETI和PTI相关,两者相互联系并在Pi9介导的稻瘟病抗性中发挥作用。【结论】与日本晴比较,抗病基因型NPB/Pi9对稻瘟病防御反应更强烈。转录因子、激酶、NBS-LRR基因、几丁质酶、水杨酸、茉莉酸和乙烯信号途径,以及植物次生代谢在Pi9介导的稻瘟病抗病反应中发挥重要作用。     

A型流感病毒分型基因芯片在流感病毒各亚型监测中的应用

为验证自行研制的A型流感病毒分型基因芯片方法在实际中的应用效果,了解A型流感病毒各亚型在我国不同地区和不同宿主中的分布情况,对我国20个地区、不同宿主源(鸡、鸭、鹌鹑、鸽子、猪、人等)拭子和组织样品共16 852份进行了检测。利用自行研制的A型流感病毒分型基因芯片方法,对样品进行RNA提取、多重不对称RT-PCR扩增、芯片杂交、洗涤和扫描分析;同时将基因芯片检测阳性的样品,用普通RT-PCR扩增后进行序列测定。结果显示:利用该方法检出阳性样品2 582份,总检出率为15.32%(2 582/16 852),共检测到H1N1、H2N9、H3N2、H4N6、H5N1、H7N1、H7N9、H9N2、H10N5、H16N3等10个亚型,且均与阳性样品测序结果一致。结果表明,该方法可准确检测不同地区、不同宿主中的A型流感病毒不同亚型。同时,该方法能在同一张芯片上准确鉴定A型流感病毒的多种亚型,解决了其他常规方法无法检测已发现和可能发生变异的所有亚型的棘手问题,从而为A型流感诊断和分子流行病学监测提供了一种新技术。     

长江流域棉花减量简化一次性施肥技术研究

针对棉花常规施肥技术存在的施肥次数多、技术复杂、用量大、利用率低、中后期土壤追肥操作不便、施肥易受天气影响等问题，研究了主栽棉花品种营养特性，研制了易降解棉花专用缓控释肥缓控释包膜新材料、系列配方，创新了生产工艺，研制的专用缓控释肥实现了棉花减量简化一次性施肥，变革了棉花施肥技术，解决了常规技术中施肥集中期与干旱雨涝等灾害高发期重叠导致的无法追肥或肥害、从而减产的重大难题。与等养分常规施肥相比，应用该技术棉花单产增加 3.8%～14.1%、增效10.1%～25.5%；减少施肥用工50%以上，等产量情况下单位面积减少施肥11.2%～52.3%。以棉花减量简化一次性施肥为核心技术，建立了棉花绿色轻简高效栽培技术体系。     

Development and Application of Cotton InDel Markers Based on High Throughput Sequencing

[Objective] Dimorphic InDel markers can be used for cotton variety identification and purity detection, to improve the accuracy and efficiency of cotton seed testing, and to play a role in molecular breeding of cotton. [Method] Based on the whole genome sequencing of 121 cotton varieties from different sources, the InDel markers with high polymorphism were developed according to polymorphism information content(PIC) and were applied in the genetic distance analysis and cluster analysis by using 66 cotton varieties in China. [Result] Totally 10 967 InDel were identified based on the next generation sequencing data of 121 cotton varieties. Among the 85 pairs of InDel primers synthesized, 64 were selected including 35 from At group and 29 from Dt group. The minimum average allelic frequency(MAF) of At and Dt chromosomes were 0.45 and 0.32, respectively, while the PIC were 0.49 and 0.40, respectively. The genetic distances of the 66 cotton varieties ranged from 0.04 to 0.65 centimorgan (cM), with an average of 0.39 cM. The two varieties with the largest genetic distance were Simian 3 and CCRI 36, and the two varieties with the smallest genetic distance were Xumian 18 and Xuza 3. [Conclusion] The 64 cotton dimorphic InDel markers can effectively reveal the relationships among varieties based on the genetic distance, and distinguish cotton varieties from different sources, which has certain theoretical significance and application value.     

山西云顶山亚高山草甸优势种种间关系分析

为揭示山西云顶山亚高山草甸植物群落优势种的种间关系,本研究通过种间联结性和相关性测定,对其植物群落13个优势种78个种对的种间关系进行研究。结果表明:13个优势种间大部分种对的种间关联不显著,种间联结不紧密,说明山西云顶山亚高山草甸植物群落结构不很稳定,存在一定程度的生态退化风险;多数原生优势种在群落中占据主要位置,同时也出现少数非原生优势种优势明显,有逐渐取代典型原生优势种的逆行演替趋势,这是由于人类频繁的旅游扰动所致。因此,应当合理的控制旅游开发强度与游客数量,保持亚高山草甸生态系统的稳定与自我维持,以持续高效的维持云顶山亚高山草甸的经济及生态价值。     

基于Kjeldahl与Dumas方法的农作物秸秆总氮含量分析

凯氏定氮法(Kjeldahl)与杜马斯燃烧法(Dumas)是测定农业生物质总氮含量的主要检测手段,但二者的测定结果数值存在差异。该研究获取农作物秸秆样本(水稻、小麦、玉米、油菜和棉花)共计1 179个,分别采用Kjeldahl和Dumas方法测定总氮(TKN和TCN,total Kjeldahl nitrogen and total combustion nitroyen)含量,通过多种统计与分析方法,系统分析比较了不同农作物秸秆总氮含量及其分布的异同和相关关系。结果表明:不同农作物秸秆氮含量分布均呈非正态分布,建议采用中位数统计;5种秸秆总体的TKN质量分数为(7.12±1.87) g/kg,TCN质量分数为(8.00±2.13) g/kg,TKN含量显著小于TCN含量;小麦和棉花秸秆的TKN含量和TCN含量与其他秸秆间均存在显著差异(P <0.05);不同生物质TKN含量与TCN含量关系不同,建议采用最小中位数二乘法进行拟合分析。研究结果可为农作物秸秆科学利用提供数据及方法互通性支撑。