植物根结线虫基因组学研究进展

根结线虫是世界农业生产中危害最大的植物病原之一,目前仍缺乏安全有效的防治措施。深入揭示寄生线虫与植物之间互作的分子机制,利用生物技术进行抗性育种被认为是最有前景的抗线虫策略。在根结线虫基因组学研究方面,目前已经构建了北方根结线虫AFLP遗传连锁图谱,南方根结线虫和北方根结线虫基因组测序也已完成;基因组的注释和比较基因组学分析,较全面地描述了根结线虫的遗传组成;以差异表达分析和比较基因组学为主的方法鉴定了大量的重要基因;以RNA干扰、植物转化和蛋白互作为主的根结线虫基因功能研究也取得了一些进展。本文就根结线虫基因组学研究予以综述,并进一步探讨其研究方向和可持续抗线虫新策略的发展前景。     

糖苷水解酶家族基因AghyGH1a参与莲草直胸跳甲对皂苷的代谢

为了解植物化合防御物质皂苷对专食性天敌莲草直胸跳甲Agasicles hygrophila的防御作用,测定莲草直胸跳甲2龄幼虫取食皂苷后的排泄和肠道变化,利用转录组测序筛选参与皂苷代谢的候选基因,并采用 RNA 干扰技术鉴定候选基因功能。结果发现,取食皂苷导致莲草直胸跳甲幼虫排泄量增加,肠道畸变,排泄量与取食皂苷浓度正相关。转录组测序发现,与对照相比,取食 100 mmol/L皂苷处理的莲草直胸跳甲幼虫肠道有496条差异表达基因,其中261个上调基因和235个下调基因。GO分类和KEGG分类分析发现糖苷水解酶家族1（glycoside hydrolase family1,GH1）基因集中在碳水化合物水解通路。利用RNA干扰技术鉴定发现,莲草直胸跳甲AghyGH1a基因沉默可导致幼虫取食皂苷后排泄量增加,肠道内容物颜色更暗沉。表明莲草直胸跳甲AghyGH1a参与皂苷代谢。     

南京中山陵风景区旅游干扰调查

随着旅游业的兴起，景区环境问题越来越得到人们的关注。以中山陵风景区为例，重点研究了旅游对中山陵风景区生态环境的影响，结果表明：该地区旅游影响了乔木的生长力（包括胸径、树高、生长缺陷），严重影响灌木、草本植物的物种多样性，其中对于高大乔木影响相对较小。提出了中山陵景区仍需要建立更为完善的保护、管理机制，维护生态稳定的建议。     

干扰水稻瘤矮病毒(RGDV)非结构蛋白(Pns12)的表达抑制病毒在介体昆虫培养细胞内的复制

水稻瘤矮病毒(Rice gall dwarf virus,RGDV)属呼肠孤病毒科(Reoviridae)植物呼肠孤病毒属(Phytoreovirus),由介体电光叶蝉(Recilia dorsalis)以持久增殖型方式传播,其基因组第12条片段编码的非结构蛋白(nonstructural protein,Pns12)是提供病毒复制和子代病毒粒体装配场所——病毒原质(viroplasm)的组分之一。为了明确Pns12在RGDV侵染介体电光叶蝉培养细胞中的功能,本研究通过原核表达的Pns12蛋白免疫注射兔(Oryctolagus cuniculus),制备Pns12抗体,并应用免疫荧光标记和RNA干扰(RNA interference,RNAi)技术研究Pns12在介体培养细胞内的定位和参与病毒原质形成的过程。共聚焦显微镜观察到,病毒侵染的细胞中,与荧光素交联的Pns12抗体特异地标记在病毒原质上。干扰Pns12蛋白表达后,可有效地阻碍病毒原质的形成、子代病毒粒体的组装和病毒非结构蛋白Pns12和外壳蛋白P8蛋白的表达,表明Pns12作为病毒原质的组分参与了RGDV在介体培养细胞内的复制。也表明,Pns12可作为理想的靶标用于阻断电光叶蝉携带和传播RGDV。     

香蕉乙烯受体基因RNA干扰表达载体的构建

设汁两对引物，PCR扩增香蕉乙烯受体基因cDNA序列自AUG启始密码子起长538bp区段的正向(记为5‘I)、反向(记为AI’)DNA区段。构建含该正、反向互补重复序列DNA片段的中间载体，经测序证实连接正确后。克隆到植物表达载体pCAMBIA-1301中的GUs基因位置并经双酶切证实。在所得表达载体pCAMBIA-1301-S5’I-AI’中，该插入正反互补重复片段处于35S启动子之后，由此转录的mRNA因两端序列反向互补而形成发夹式RNA，因此可应用于RNA干扰研究。同时，文中对构建含正、反向重复序列插入片段植物表达载体过程中出现的插入片段链内退火引起连接困难等问题进行了分析讨论。     

小麦种群邻体干扰模型的研究

改进的邻体干扰模型为：Ｙ＝Ｙｍ（１＋ＣＷ＾т）ˉ１。其中，Ｙｍ为不存在邻体时的最大生物量，Ｃ，Ｔ为常数。     

拟小食螨瓢虫成虫对朱砂叶螨捕食作用的研究

拟小食螨瓢虫是一种栖息于木薯上的朱砂叶螨的重要天敌。其成虫能捕食朱砂叶螨的各个虫态,随着猎物密度的增加其捕食量逐渐上升,对朱砂叶螨卵、幼螨、若螨和成螨的日捕食量上限分别为146粒,87、91和11头。当猎物密度不变时,随着拟小食螨瓢虫成虫密度的增加,其捕食量下降。     

RNA干扰抑制ST细胞Ⅰ型干扰素受体-2表达的研究

应用RNA干扰技术建立猪睾丸(ST)细胞Ⅰ型干扰素受体-2(IFNAR-2)基因knock down模型。采用FuGENE HD转染剂将pSiCMVE1、pSiCMVE2和pSiCMVE3干扰载体分别转染ST细胞,经荧光定量PCR和Western blot分别检测IFNAR2mRNA和蛋白水平表达变化。结果显示,与对照组比,转染干扰载体组均可明显降低IFNAR2 mRNA和蛋白的表达(P0.05)。RNA干扰技术能够有效抑制IFNAR2基因的表达,为进一步研究IFNAR2基因的生物学功能提供可靠的手段,为研究IFNAR2在病毒感染中的作用奠定基础。     

Study on the Effect of Infectious Bronchitis Virus and Newcastle Disease Virus on Proliferation and Immune Interference

This experiment aimed to investigate the proliferation interference effects of the infectious bronchitis virus (IBV) and Newcastle disease virus (NDV),analyze the immune interference of two vaccines in the immune process,and provide a basis for determining vaccine immune program.This experiment was in a complete randomized design,10-day-old allantoic cavity of the chick embryo was inoculated with different concentrations of IBV and NDV and with a mixture of different concentrations of IBV and NDV,and then the allantoic fluid was collected for determination of the titer of hemagglutination (HA).Furthermore,BALB/c mice and SPF chicken were immunized with different concentrations of IBV and NDV and also with the mixed IBV and NDV with different concentrations,the blood of mice and chicken were collected,and antibody titer and inhibitory titer of IBV and NDV were determined by indirect enzyme linked immunosorbent assay (ELISA) and hemagglutination inhibition (HI) test.Experimental results showed that in the homeomorphic cultivation research,whether IBV and NDV were inoculated in a different order or inoculated at the same time,the inoculation of IBV behaved interference to the NDV,while the inoculation of NDV had no interference effects to IBV;Mice were inoculated with IBV and NDV,IBV and NDV antibody titers were lower than single immunized group,immune procedures and immune intervals often had effect on the titer of IBV and NDV antibody,IBV had interference on the immune effect of NDV,different needle mixed immunization can improve the antibody titer of IBV and NDV.In the group of SPF chicken inoculated with IBV and NDV,the order of the inoculation and the immune interval had effect on the titer of IBV and NDV antibody,IBV had a interference on the immune effect of NDV;In the immune serum inhibitory test,the data showed that the IBV and NDV antibody production were affected by the immune order of the inoculation and the interval,the effect on the NDV antibody was more obvious,but with the increase of the immune interval,the antibody level of IBV and NDV showed an increasing trend,the chicken was more sensitive than the mice to reflect the immune interference of IBV on NDV.IBVand NDV antibodies titer was also reduced in the groups that was inoculated with the mixed IBV and NDV,IBV had a interference on the immune effect of NDV.In the homeomorphic cultivation research and animal research,IBV behaved interference to the NDV,while NDV had no interference effects to IBV.IBV and NDV antibody titers decreased when mixed immunization,IBV had a interference on the immune effect of NDV.The order of the inoculation and the immune interval had effect on the antibody titer of IBV and NDV,but with the increase of the immune interval,the antibody level of IBV and NDV showed an increasing trend.     

“回避行为”初探

心理语言学日新月异的发展推动着“错误分析”理论由外显的,线性平面上的干扰研究发展到内在的,深层的非直接性干扰研究。根据这种发展动态,本文对“回避行为”的假设先决条件与思想动因作了较为深刻的探讨与剖析。