高通量测序发现河南开封、中牟西瓜病毒病由多种病毒复合侵染导致

 调查发现河南省开封市祥符区和郑州市中牟县西瓜田病毒病严重发生,症状表现为叶片黄化、蕨叶、皱缩、卷曲,枝梢翘起,果实变小和瓜纹不清。为明确其病毒种类,通过小RNA测序及生物信息学分析发现采集的西瓜样本中存在8种病毒。其中包括5种已知病毒:小西葫芦黄花叶病毒(Zucchini yellow mosaic virus, ZYMV)、甜瓜蚜传黄化病毒(Melon aphid-borne yellows virus, MABYV)、西瓜花叶病毒(Watermelon mosaic virus, WMV)、黄瓜绿斑驳花叶病毒(Cucumber green mottle mo-saic virus, CGMMV)和西瓜隐潜病毒(Citrullus lanatus cryptic virus, CiLCV);3种为新检出的RNA病毒:西瓜病毒A(Watermelon virus A, WVA)、西瓜皱叶相关病毒1号(Watermelon crinkle leaf-associated virus 1, WCLaV-1)和西瓜皱叶相关病毒2号(Watermelon crinkle leaf-associated virus 2, WCLaV-2)。RT-PCR检测田间66个西瓜标样发现,WMV、CiLCV和ZYMV是侵染两地西瓜的主要病毒,其阳性检出率分别为61%、56%和52%,两种及以上病毒复合侵染的比率达68%。     

白背飞虱酚氧化酶原PPO基因特性及其免疫应答

【目的】酚氧化酶（phenoloxidase,PO）是昆虫体内的免疫蛋白,在昆虫体内起重要的调节作用,主要以无活性的酚氧化酶原（prophenoloxidase,PPO）形式存在。本研究在转录组测序获得白背飞虱（Sogatella furcifera）3条PPO序列的基础上,通过分析白背飞虱体内不同SfPPO的发育和组织表达模式,并进一步抑制SfPPO的表达以及病原菌诱导,探讨SfPPO在白背飞虱体内的生物学功能。【方法】以白背飞虱3条SfPPO序列为研究对象,利用生物信息学方法分析其蛋白结构以及与其他昆虫之间的同源性。并以注射绿色荧光蛋白（green fluorescent protein,GFP）的dsRNA作为对照组,通过注射合成的dsSfPPO后采用实时荧光定量PCR（qRT-PCR）检测基因表达抑制效果;此外,为了探究SfPPO在白背飞虱生长发育和免疫应答方面的作用,利用qRT-PCR检测SfPPO在不同发育阶段及组织中的表达情况,以及注射大肠杆菌（Escherichia coli）和枯草芽孢杆菌（Bacillus subtilis）后3个SfPPO的诱导表达情况。【结果】SfPPO1、SfPPO2-1、SfPPO2-2的开放阅读框长度分别为2 079、999、2 070 bp,分别编码692、332、689个氨基酸,预测蛋白分子量分别为79.84、37.67、79.53 kD,等电点分别为6.43、9.56、6.20。生物信息学分析表明,白背飞虱的3个PPO蛋白具有较高的同源性,且与褐飞虱（Nilaparvata lugens）的亲缘关系最近;SfPPO1、SfPPO2-1、SfPPO2-2均在成虫第3天表达量最高,其中SfPPO2-1和SfPPO2-2的发育表达模式相似;SfPPO1和SfPPO2-1在表皮和脂肪体内表达量较高,而SfPPO2-2在翅和脂肪体内表达量较高,在头、足、中肠、马氏管中表达量相对较低;与注射dsGFP组相比,注射靶标基因的dsRNA后都能够显著沉默目的基因的表达,而且当SfPPO2-1表达被沉默后,SfPPO1出现显著高表达,说明不同的PPO之间可能具有补偿功能;大肠杆菌诱导后12 h,SfPPO2-1和SfPPO2-2表达量显著升高,SfPPO1表达量则在诱导24 h后显著升高;枯草芽胞杆菌诱导24 h后,SfPPO1、SfPPO2-1和SfPPO2-2表达量均显著升高。【结论】SfPPO1、SfPPO2-1、SfPPO2-2的表达存在组织和龄期特异性,且在不同菌诱导情况下存在免疫应答差异性。研究结果有助于更加全面、深入地探索白背飞虱酚氧化酶在调控昆虫发育及免疫中的潜在分子机理。     

水稻种子总RNA提取质量问题的探究

水稻种子总RNA提取在水稻分子遗传基础实验中起着非常重要的作用,然而,高质量水稻种子RNA的获取难度高,极大阻碍了cDNA文库构建、RNA-Seq和基因表达分析等相关研究工作的开展和深入。为了探讨水稻种子总RNA提取的相关影响因素,寻找合适的水稻种子总RNA提取方式,本研究通过比较Trizol法、GeneMark试剂盒法和艾德莱试剂盒法3种不同的提取方式、不同的耗材处理方法、研磨方式等不同因素改进水稻种子总RNA的提取效果。不同的检测结果显示,艾德莱多糖多酚提取试剂盒能够提取到28S、18S条带完整、降解少和纯度高的水稻种子总RNA;不同耗材处理方法中,蛋白酶K处理过的耗材提取的总RNA比另外两种方法处理过的耗材提取的总RNA降解更少,28S和18S条带更清晰和完整、总RNA质量最高、效果最好;而研磨方式上,液氮冷冻研磨法能够有效抑制RNA酶对RNA的降解。本研究对水稻种子RNA提取过程相关因素的分析和改进,为促进以水稻种子为研究对象的分子实验奠定了一定的技术基础。     

全双工双通路中继网络干扰处理与信号检测算法

全双工双通路连续中继(FD-TPSR)网络频谱效率高,状态控制简单,但全双工中继存在残留自干扰(RSI),发射中继对接收中继也会形成中继间干扰(IRI),会导致通信性能下降.这里提出一种中继间干扰处理方法,中继节点消除部分IRI,以提高端到端信干噪比;保留部分IRI,在目的节点构成延迟转发编码结构,以提供分集增益.同时提出一种基于并行软干扰消除的匹配滤波(MF-PSIC)信号检测算法,匹配滤波器结构简单,实现复杂度低,基于软输出的并行干扰消除能同时检测所有时隙的符号,处理时延小.仿真结果表明,中继间干扰处理方法兼顾了分集增益和累积干扰与噪声因素,相比于无IRI消除和完全IRI消除,误比特率最低; MF-PSIC信号检测算法实现复杂度低,相比于ML信号检测算法仅有很少的性能损失.     

Performance of Bangladesh indigenous rice in a weed infested field and separation of allelopathy from resource competition

This study aimed to identify the potential allelopathic indigenous rice (Oryza sativa L. ssp. indica) varieties from Bangladesh using a performance study in a weed‐infested field and to assess the extent of allelopathic interference relative to resource competition in a glasshouse experiment. Six varieties – namely, “Boterswar,” “Goria,” “Biron” and “Kartiksail” as the most allelopathic, “Hashikolmi” as weakly allelopathic and “Holoi” as nonallelopathic – were raised following a nonweed control method. The infestation levels of weed species were calculated using Simpson's Diversity Index (SDI), which ranged from 0.2 to 0.56. However, a significant correlation coefficient (0.87, P < 0.001) was obtained from these field data compared with the root inhibition percentage from the laboratory bioassay, and the “Boterswar” variety was the most allelopathic. The interactions between the allelopathic variety “Boterswar,” weakly allelopathic variety “Hashikolmi” and Echinochloa oryzicola via a target (rice)‐adjacent (E. oryzicola) cogrowth culture were determined in a hydroponic arrangement. The relative competitive intensity (RCI) and the relative neighbor effect (RNE) values showed that the crop–weed interaction was facilitation for “Boterswar” and competition for “Hashikolmi” and E. oryzicola in rice/E. oryzicola cogrowth cultures. The allelopathic effects of “Boterswar” were much higher than the resource competition in rice/E. oryzicola cogrowth cultures. The converse was observed for “Hashikolmi.” Moreover, the mineral content of E. oryzicola was severely affected by “Boterswar”/E. oryzicola cogrowth cultures’ exudate solution. Therefore, the allelopathic potential of “Boterswar” variety might be useful for developing the weed‐suppressing capacity of rice, which will likely have a significant influence on paddy weed control.     

Putting the pieces together: Recent growth,nutritional condition,and mortality of Engraulis anchoita larvae in the Southwest Atlantic

Dynamics of clupeiform fish populations such as anchovy are frequently impacted by environmental variations which can affect the success of the species recruitment. Herein, we have analyzed recent otolith growth rate, RNA/DNA nutritional condition index (sRD), and mortality rate of argentine anchovy larvae Engraulis anchoita from three different nursery areas in the Southwest Atlantic. We have evaluated the relationship between the environmental variables (abundance of copepod nauplii, temperature, chlorophyll‐a concentration, and abundance of E. anchoita larvae) and larval endogenous variables (size, weight, age, and otolith radius) to sRD and recent growth rate. Fast larval growth rates were observed toward the northern sector of the studied area, characterized by higher temperature. High values of sRD were associated with higher nauplii abundance in the proximity of coastal fronts. The larvae with the lowest growths and lowest minimum values of nutritional condition coincided with the area where there was less abundance of nauplii and higher larval mortality. Larval size and nauplii abundance were positive explanatory factors for both recent growth rate and sRD index. Temperature had a positive effect on recent growth rate and a negative effect on sRD index. This condition index was poorly explained in terms of model fit in comparison with the growth model. The results herein provided could be significant to better understand the recruitment of the species, as to determining favorable areas for the growth and survival of anchovy larvae.     

寄主诱导的基因沉默提高植物真菌病害抗性研究进展

寄主诱导的基因沉默(host-induced gene silencing,HIGS)以病原菌生长发育、产孢繁殖、侵染或致病过程中的关键基因作为靶点,在寄主植物中表达针对靶基因的RNAi构建体,在病原菌侵染植物的过程中,摄入相应的ds RNA或si RNA分子,通过识别、结合特异核苷酸序列,干扰病菌靶基因表达,从而抑制病菌侵染和扩展,使植物表现抗病。这项技术为培育基于病原菌特异序列的植物抗病性奠定了基础,显示了巨大的应用潜力。本文综述了利用HIGS技术提高植物真菌病抗性的最新研究进展,总结了国内外利用这项技术改良植物真菌病害抗性的主要策略、技术路线,展望了发展应用前景。     

试验干扰下土壤中微量元素空间变异性研究

[目的]为了解烟草试验站土壤中微量元素的空间变异性,进而为大田试验布局提供依据。[方法]用地统计学方法,分析彭水试验站的耕层土壤养分的空间变异特征。[结果]该站内土壤有效铁、有效铜、有效钙的含量丰富;有效锌和有效锰为中等含量水平,有效硫和有效镁含量从低至高均有分布。各中微量元素间具有显著的相关性。所有元素均可用指数模型和球状模型进行较好拟合;有效锌、有效镁和有效硫由于受结构性因素影响而呈现强空间自相关性;其余元素的空间自相关性由结构性因素和随机因素共同作用而呈现中等空间相关性。Kriging插值显示,东部及东北部较高,中部和西南部区域含量较低;有效铁、有效钙和有效硫的空间分布的规律性差。[结论]该试验站土壤中微量元素含量存在较大的空间变异性。在安排大田试验时,要充分认识这种变异性,合理布局大田试验。     

油菜种子低亚油酸和亚麻酸含量异交稳定技术的建立

为降低油菜种子中的亚油酸和亚麻酸含量并使之在异交条件下保持基本稳定,以高油品种CY2作为受体亲本,采用RNA干涉技术沉默油菜内源Bn FAD2基因的表达,抑制油酸脱饱和反应。育成的3个独立转基因株系的亚油酸和亚麻酸含量下降到6%以下,其中亚油酸含量较受体亲本降低78.2%~86.5%,亚麻酸含量降低53.4%~65.8%。将3个转基因株系与2个亚油酸和亚麻酸含量较高的常规品种正反交,F1种子中这2种脂肪酸的含量依然保持在与转基因亲本相仿的低水平。由此可知,通过RNA干涉技术沉默Bn FAD2基因的表达可赋予油菜低亚油酸和亚麻酸含量的异交稳定特性。这一特性的获得对于今后培育异交稳定的高油酸油菜新品种具有重要意义。