Virus transmission by aphids in potato crops

This paper reviews the contribution of vector activity and plant age to virus spread in potato crops. Determining which aphid species are vectors is particularly important for timing haulm destruction to minimize tuber infection by potato virus Y (PVY). Alate aphids of more than 30 species transmit PVY, and aphids such asRhopalosiphum padi, that migrate in large numbers before flights of the more efficient vector,Myzus persicae, appear to be important vectors. Differences in methodology, aphid biotypes and virus strains prevent direct comparisons between estimates of vector efficiencies obtained for aphids in different countries in north western Europe. M. persicae is also the most efficient vector of potato leafroll virus (PLRV), but some clones ofMacrosiphum euphorbiae transmit PLRV efficiently toNicotiana clevelandii and potato test plants. The removal of infected plants early in the season prevents the spread of PLRV in cool regions with limited vector activity. The proportion of aphids acquiring PLRV from infected potato plants decreases with plant age, and healthy potato plants are more resistant to infection later in the season. Severe symptoms of secondary leafroll developed on progeny plants of cv. Maris Piper derived from mother plants inoculated with PLRV in June or July of the previous year. Progeny plants derived from mother plants inoculated in August showed only mild symptoms, but the concentration of PLRV in these plants was as high as that in the plants with severe symptoms.     

马铃薯卷叶病毒PLRV RT-LAMP检测方法优化

马铃薯卷叶病毒Potato leafroll virus(PLRV)是目前严重影响马铃薯产量与品质的主要病毒之一,给马铃薯产业造成巨大损失。本研究采用环介导等温核酸扩增(loop-mediated isothermal amplification, LAMP)技术建立PLRV的RT-LAMP检测方法。采取单因素变化试验,对RT-LAMP反应体系中多个因素包括引物组合、温度条件及Mg~(2+)、betaine、Bst 3.0 DNA聚合酶、dNTPs、UNG、SYBR GreenⅠ和引物组合的浓度进行一系列试验和优化。采用RT-PCR检测方法进行平行比对试验,对优化后的RT-LAMP反应体系进行了验证。结果表明,最佳引物组合为P3,最适反应温度62℃,25μL反应体系中,Mg~(2+)、betaine、Bst 3.0 DNA聚合酶和UNG的最佳终浓度分别为4 mmol/L、0 mmol/L、0.64 U/μL和0.08 U/μL,dNTPs的最佳用量为1μL(dATP、dGTP、dCTP各0.4 mmol/L,dUTP 1.2 mmol/L),SYBR GreenⅠ(20×)的最佳用量1μL,primer mix的最佳用量2.5μL(PLRV-FIP/BIP、PLRV-F3/B3和PLRV-LF/LB的浓度分别为0.8、0.2μmol/L和0.6μmol/L),RNA模板1μL(2 ng/μL),加DEPC-H_2O至25μL,反应时间50 min。优化后的RT-LAMP检测结果与RT-PCR一致,且可视化判读结果。因此,建立的PLRV RT-LAMP检测方法为进一步开发RT-LAMP检测试剂盒及其实际应用奠定了基础。     

马铃薯卷叶病毒的RT-PCR快速检测

根据马铃薯卷叶病毒的外壳蛋白基因序列,设计合成了一对寡核苷酸引物。从感染马铃薯卷叶病毒(PLRV)的马铃薯叶片中提取出病毒RNA,进行cDNA合成并运用RT-PCR技术进行体外扩增,得到一条长度约627bp的特异PCR扩增产物,与理论设计的外壳蛋白基因大小一致,而对照未得到任何产物。从而建立了快速灵敏的PLRV检测方法,为PLRV的防治及检测提供了有效手段。     

An antisense coat protein gene confers immunity to potato leafroll virus in a genetically engineered potato

The Bzura commercial potato cultivar was transformed by sense or antisense constructs which included the coat protein gene of potato leafroll virus RNA. In the sense construct, the coat protein gene was preceded by a leader sequence shorter than that in the subgenomic RNA formed in infected cells. The antisense construct consisted of a sequence complementary to the first 2020 nucleotides of the subgenomic RNA. Selected transformants expressing viral RNA were resistant to virus challenge by viruliferous aphids. In one line, expression of the antisense RNA prevented virus infection even after grafting with scions from infected plants and therefore this transformant might be regarded as virus immune.     

利用TPS夏播生产优质实生种薯的研究──夏播实生种薯和常规实生种薯的比较

通过夏播实生种薯和常规实生种薯的比较试验，揭示了夏播实生种薯具有带毒少、出苗率高、长势强、退化轻、晚疫病轻、产量和淀粉含量明显提高等特点．进一步证明利用马铃薯实生种子夏播生产优质实生种薯是效果显著，完全可行的。     

马铃薯PVY和PLRV病毒的TC-RT-PCR检测

传统的RT-PCR技术检测病毒需提取总RNA,RNA容易降解。利用试管捕捉反转录扩增(Tube cap-ture RT-PCR,TC-RT-PCR)方法检测了PVY和PLRV 2种病毒,实现了不需提取总RNA也可在同一反应中同时检测2种病毒。根据已报道的引物用TC-RT-PCR的方法对PVY和PLRV的外壳蛋白基因进行了检测。结果表明,TC-RT-PCR能够成功的检测出感染PVY或PLRV以及2种病毒共同侵染的样品,扩增产物序列长度均与设计片段的长度相符,分别为781和364 bp,2种病毒扩增产物的测序结果同Gene Bank中已知的序列比对后的同源性均高达97%以上。该技术为单独或复合感染的马铃薯病毒的检测提供了更加方便、高效的方法。同时测得试验PVY病毒样本属于PVYNW株系。     

Argentinian potato leafroll virus P0 protein: Novel activities for a previously known suppressor

The potato leafroll virus (PLRV) P0 protein (P0^PL) is a suppressor of RNA silencing. In this study, we showed that P0 protein from an Argentinian isolate of PLRV (P0^PL-Ar) has an additional activity not described for other PLRV or P0 proteins from poleroviruses. Besides reporting that P0^PL-Ar displays both local and systemic silencing suppressor activity, we demonstrated, for the first time, that P0^PL-Ar impedes accumulation of dsRNA-derived siRNAs. We also showed that P0^PL-Ar interacts with Solanum tuberosum SKP1 orthologue (StSKP1) and triggers destabilization of ARGONAUTE 1 (AGO1) and that these actions are mediated by the F-box-like domain. A mutant in the GW/WG motif within the P0^PL-Ar F-box-like motif lost the suppression activity, the interaction with StSKP1 and abolished AGO1 decay. Interestingly, a mutant in the L76/P77 residues within the P0^PL-Ar F-box-like motif, which lost the suppression activity and the interaction with StSKP1, retained the capacity to enable AGO1 decay. Thus, unlike other P0 proteins of previously characterized poleroviruses, P0^PL-Ar seems to have a dual activity, according to the findings of this study. This protein would act at both an upstream and a downstream step of the RNA silencing pathway: upstream of Dicer-like enzyme (DCL)-mediated primary siRNA production and downstream at the RNA-induced silencing complex (RISC) complex level. Our results contribute to the understanding of the different ways PLRV P0 proteins function as silencing suppressors.     

Evidence that resistance to potato leafroll virus accumulation in tetraploidsolarium tuberosum L. is controlled by one or few major genes that are not complementary

Summary Previous investigations into the inheritance of resistance to accumulation of potato leafroll virus indicated a dominant major-gene effect, but the segregation ratios in progenies of crosses were a closer fit to a model involving two complementary genes (both required for resistance and one present in the susceptible parent cultivar, Maris Piper) than to a single gene model. In this study, we tested the complementary gene hypothesis by backcrossing susceptible segregants from one of these progenies to their susceptible parent, Maris Piper. No resistant segregants were found in the five backcross progenies examined, so the complementary gene hypothesis was not supported. There was significant variation between susceptible progeny-members in these backcrosses. The progeny of another, highly resistant parent clone, G. 8107(1), selfed, was also examined: all members were resistant. Whilst there is evidently a dominant major-gene effect involved, this is not the whole picture and there are other unidentified genetic effects.     

马铃薯不同级别脱毒种薯病毒再侵染情况及产量变化

通过对大西洋原原种、一级原种、二级原种、一级种薯和二级种薯的生产试验及其PVX、PVY、PVS、PLRV病毒的DAS-ELISA检测,结果表明,从原原种到一级种薯的生产过程中,产量随着脱毒种薯代数的增加而升高,其中以二级原种生产一级种薯的增产幅度最显著;除原原种不带以上4种病毒外,一级原种、二级原种分别带3%、13.3%的PVS病毒,一级种薯带PLRV和PVS病毒,带毒率分别是5.8%、21%,二级种薯带PVY、PVS、PLRV病毒,带毒率分别是8.9%、30%、13%.