华山松SSR-PCR反应体系优化

[目的]建立华山松SSR-PCR最佳反应体系,为华山松种质资源的分子标记辅助育种、遗传多样性分析及遗传图谱构建研究提供理论参考.[方法]以华山松幼嫩针叶为材料,分别采用试剂盒法、SDS法和改良CTAB法提取华山松基因组DNA,确定华山松DNA最佳的提取方法.通过L16(44)正交试验设计对华山松SSR-PCR反应体系中引物用量(A)、dNTP用量(B)、Taq DNA聚合酶用量(C)和DNA模板用量(D)进行优化,获得华山松SSR-PCR最佳反应体系.[结果]改良CTAB法提取的基因组DNA浓度为92.1~1786.3 ng/μL,OD260/OD280为1.80~2.07,OD260/OD230比值约2.0,条带明亮且清晰,无弥散现象,表明该方法提取效果最佳.正交试验极差分析结果显示,4个因素影响华山松SSR-PCR反应体系扩增效果的主次顺序为A>D>B>C,最优水平组合为A4B2C2D2.正交试验方差分析结果显示,4个因素影响华山松SSR-PCR反应体系扩增效果的主次顺序为A>D>C>B,最优水平组合为A4D2C2B2.结合正交试验16个组合的评分结果,最终确定华山松SSR-PCR最佳反应体系(20.00μL):10μmol/L引物0.35μL,50 ng/μL DNA模板1.00μL,10 mmol/L dNTP 2.00μL,5 U/μL Taq DNA聚合酶1.20μL,10×PCR Buffer(含Mg2+15 mmol/L)2.00μL,用ddH2O补足至20.00μL.[结论]优化获得的华山松SSR-PCR最佳反应体系可应用于华山松种质资源评价及分子标记辅助育种等研究.     

Simulation modelling of potato virus Y spread in relation to initial inoculum and vector activity

Potato virus Y(PVY) is a non-persistent virus that is transmitted by many aphid species and causes significant damage to potato production. We constructed a spatially-explicit model simulating PVY spread in a potato field and used it to investigate possible effects of transmission efficiency, initial inoculum levels, vector behavior, vector abundance, and timing of peak vector activity on PVY incidence at the end of a simulated growing season. Lower PVY incidence in planted seed resulted in lower virus infection at the end of the season. However, when populations of efficient PVY vectors were high, significant PVY spread occurred even when initial virus inoculum was low. Non-colonizing aphids were more important for PVY spread compared to colonizing aphids, particularly at high densities. An early-season peak in the numbers of noncolonizing aphids resulted in the highest number of infected plants in the end of the season, while mid-and late-season peaks caused relatively little virus spread. Our results highlight the importance of integrating different techniques to prevent the number of PVY-infected plants from exceeding economically acceptable levels instead of trying to control aphids within potato fields. Such management plans should be implemented very early in a growing season.     

热带果蔬采后冷害研究进展

果蔬采后冷害机制及其防控是当前采后研究领域中倍受关注的科技问题。果蔬冷害发生是由内在因子和外部环境因素共同作用的结果。近年来,随着现代分子生物学技术的应用,并借鉴模式植物拟南芥的研究成果,果蔬采后冷害发生调控机理及防控技术研究取得一系列进展。本文综述了热带果蔬采后冷害发生的生理与分子机制和冷害防控技术的最新研究成果,关注能量代谢调控果蔬采后冷害作用,重点介绍了生物技术在防控果蔬冷害上的应用,并对今后研究方向作了展望。     

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。     

“金花散茶”及“金花菌粉”对被动吸烟小鼠肺组织JAK2/STAT3炎性及磷酸化蛋白表达的影响

为探究“金花散茶”及其“金花菌粉”对被动吸烟(Cigarette smoking environment,CSE)小鼠肺组织受损的预防及修复机制,建立C57BL/6小鼠CSE模型,以600 mg∙kg^-1剂量的金花散茶茶汤(Eurotium cristatum tea extract,ECTE)及金花菌粉浸提液(Eurotium cristatum powder extract,ECPE)进行灌喂处理。与CSE模型组相比,小鼠灌喂ECPE和ECTE后,肺组织病理学切片显示其可保护小鼠肺组织形态结构完整;酶联免疫分析显示,灌喂ECPE和ECTE可显著抑制小鼠血清IL-6、IL-8、IL-1β、IFN-γ和TNF-α表达量上调;Western blot结果表明,灌喂ECPE和ECTE对小鼠肺组织p-JAK2、p-STAT3、p-JAK2/JAK2、p-STAT3/STAT3高表达起到抑制作用。以上研究结果表明,灌喂ECPE、ECTE对CSE肺受损小鼠具有明显保护作用,总体趋势为ECPE组优于ECTE组、预防组优于治疗组。     

外源5-氨基乙酰丙酸对低温胁迫下茶树叶片光合及生理特性的影响

通过对陕茶1号（耐低温型）和金牡丹（低温敏感型）2个茶树品种在冬季自然低温胁迫下喷施不同浓度（0、10、30、50 mg·L^-1）的外源5-氨基乙酰丙酸（5-aminolevulinic acid,ALA）,探究外源ALA对低温胁迫下茶树叶片光合荧光特性及生理特性的调控作用。结果表明,适宜浓度的外源ALA对低温胁迫下茶树叶片净光合速率、气孔导度、胞间CO₂浓度、蒸腾速率、PSⅡ最大光化学效率及PSⅡ潜在活性具有促进作用,能够提高水浸出物、咖啡碱、游离氨基酸、儿茶素类物质、茶氨酸、可溶性糖、可溶性蛋白等生理活性物质含量的累积;外源ALA能够提高低温胁迫下茶树光合作用的能力并改善茶叶品质,其中50 mg·L^-1的ALA处理能有效提高陕茶1号应对低温胁迫的能力,10 mg·L^-1和30 mg·L^-1的外源ALA对金牡丹应对低温胁迫具有较好的缓解效用。     

2019年美国玉米高产竞赛简报

介绍2019年美国玉米高产竞赛情况和竞赛结果,分析全美优胜者的分布区域、选用种子品牌及信息,进而分析玉米高产原因,从而提出美国玉米高产竞赛对我国玉米生产的启示和建议。     

Effects of platelet-rich plasma on chondrocyte apoptosis and NLRP3/IL-1β signaling pathway in rabbits with osteoarthritis

AIM To explore the effect of platelet-rich plasma (PRP) on rabbit osteoarthritis and its possible mechanism. METHODS The rabbits with knee osteoarthritis were prepared and then divided into model group, sodium hyaluronate (SH) group and PRP group, and another sham operation group was set up, with 6 rabbits in each group. The gross morphological changes of rabbit cartilage were observed. HE staining was used to evaluate the pathomorphological changes of the cartilage. TUNEL staining was used to detect the apoptosis of chondrocytes. The expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/interleukin-1β （IL-1β） signaling pathway-related molecules was observed by immunohistochemical staining, and the protein levels of caspase-3, Bcl-2 and Bax were determined by Western blot. Chondrocytes were isolated and processed according to grouping, and the NLRP3 and IL-1β levels of the cells were measured by ELISA. RESULTS Compared with sham operation group, Pelletier score, Mankin score, chondrocyte apoptotic rate, the positive protein expression rates of NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), caspase-1 and IL-1β, and the protein levels of caspase-3 and Bax in model group were increased significantly (P<0.05), while the protein expression of Bcl-2 was decreased significantly (P<0.05). Compared with model group, Pelletier score, Mankin score, the apoptotic rate of chondrocytes, the positive protein expression rates of NLRP3, ASC, caspase-1 and IL-1β, and the protein levels of caspase-3 and Bax in SH group and PRP group were decreased significantly (P<0.05), while the protein expression of Bcl-2 was increased significantly (P<0.05). In PRP group, Pelletier score, Mankin score, the apoptotic rate of chondrocytes, the positive protein expression rates of NLRP3, ASC, caspase-1 and IL-1β, and the protein levels of caspase-3 and Bax were lower than those in SH group, while the protein expression of Bcl-2 was higher than that in SH group (P<0.05). Compared with control group, the expression of NL?RP3 and IL-1β in MCC950 (NLRP3 ihibitor) group were significantly reduced (P<0.05), the expression of NLRP3 in eucalyptol (IL-1β inhibitor) group was not significantly changed (P>0.05), and the expression of IL-1β was significantly reduced (P<0.05). CONCLUSION Platelet-rich plasma promotes the repair of cartilage in osteoarthritis rabbits, which has better effect than SH. The mechanism may be related to the inhibition of NLRP3/IL-1β pathway and the reduction of chondrocyte apoptosis.     

MCP-1/CCR2 signaling pathway mediates alcohol-induced angiogenesis in breast cancer

AIM To investigate the role of monocyte chemoattractant protein-1 (MCP-1) and its receptor CC chemokine receptor 2 (CCR2) in ethanol-promoted breast cancer angiogenesis and the underlying mechanism. METH?ODS: A mouse model of transplanted breast tumor with moderate alcohol consumption was established. The correlations between the expression of MCP-1/CCR2 and the expression of angiogenesis markers [platelet endothelial cell adhesion molecule-1 (PECAM-1) and vascular endothelial growth factor (VEGF)] in tumor tissues were examined by immunohistochemistry. In vitro, a 3D tumor-endothelial co-culture system was established to observe tumor angiogenesis and the role of MCP-1/CCR2 signaling pathway in alcohol-mediated angiogenesis. The cell migration ability was detected to clarify whether MCP-1/CCR2 enhanced cell mobility to form new vessels. RESULTS MCP-1 and CCR2 were both highly expressed in the breast tumor tissues of tumor-bearing mice consuming alcohol, and their expression levels were consistent with the angiogenic markers PECAM-1 and VEGF (P<0.05). The interaction between mouse breast cancer E0771 cells and endothelial cells was observed to promote angiogenesis in the 3D tumor-endothelial co-culture system with or without alcohol stimulation. MCP-1 promoted this kind of tumor angiogenesis, while CCR2 antagonist effectively inhibited the tumor angiogenesis and especially blocked alcohol-induced angiogenesis. Activation of MCP-1/CCR2 signaling pathway enhanced the migration ability of endothelial cells. CONCLUSION The MCP-1/CCR2 signaling pathway plays an important role in promoting the angiogenesis of breast cancer stimulated by alcohol. The mechanism might be that MCP-1 improves the migration of endothelial cells and then promotes angiogenesis.     

甜瓜嫩果皮颜色遗传规律研究

为探究甜瓜嫩果皮颜色的遗传规律,以甜瓜嫩果皮颜色为深绿色的S26和青色的S28为亲本材料,通过构建BC1群体,利用卡方测验,分析回交群体BC1的深绿色嫩果皮和青色嫩果皮的分离比例,开展甜瓜嫩果皮的遗传规律分析,以对嫩果皮颜色基因进行初步定位。结果表明：BC1群体表型表现为深绿色和青色2种,且群体比例为1∶1,从而确定颜色性状为单基因控制且深绿色为显性性状。通过分离群体分组分析法（BSA）和混池测序结果可以看出,控制甜瓜嫩果皮颜色的基因位于chr04的端部位置,这为后续基因的精细定位以及克隆提供了研究基础。