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1.
马铃薯Y病毒一步RT-PCR检测试剂盒的研制 总被引：1，自引：0，他引：1
马铃薯Y病毒(Potato virus Y,PVY)对马铃薯的危害最大,可导致马铃薯退化,降低马铃薯产量。解决这一问题的重要途径就是培养脱毒种薯,但是否完全脱毒需要经过检测才能证实。本研究依据PVY CP基因序列设计合成了一对引物PY1、PY2,以带毒样品植物总RNA为模板,在同一个反应中同时加入反转录和PCR反应所需试剂,反应程序中包括反转录和PCR反应所需条件,进行反应扩增,带毒样品扩增得到340 bp的目的条带,而健康对照无此目的条带,从而建立了PVY的一步RT-PCR检测技术,并组装成试剂盒。该试剂盒具有良好的稳定性和特异性,灵敏度可以检测到带毒植物组织下限的6.25μg,高于ELISA(100μg)和NASH(15μg)的灵敏度,虽然和常规方法的灵敏度相同,但更为快速、简便、易于操作,适合脱毒苗和脱毒种薯生产单位做大量样品的检测。 相似文献
2.
湖南省马铃薯主产区马铃薯病毒种类及流行分析 总被引：2，自引：0，他引：2
马铃薯是世界第四大粮食作物,其病毒病危害严重。2010年对湖南马铃薯主产区采集的66个病毒标样进行了RT-PCR检测,结果表明,检测出的马铃薯病毒有马铃薯Y病毒(PVY)、马铃薯卷叶病毒(PLRV)、马铃薯X病毒(PVX)、马铃薯S病毒(PVS)、马铃薯A病毒(PVA)和马铃薯纺锤块茎类病毒(PSTVd)。其中PVS的检出率最高,为54.5%,其次是PVX,检出率为45.5%,PVY的检出率为39.4%,PSTVd和PVA的检出率均为21.2%,PLRV的检出率为18.2%。2~4种病毒的复合侵染现象较为普遍。PVY中重组型PVY占85.7%。 相似文献
3.
将多种病毒的有效核酸片断拼接成融合基因转入马铃薯可获得多抗马铃薯材料。针对马铃薯生产中分布广泛、危害严重并经常混合感染的马铃薯X病毒(PVX)、马铃薯Y病毒(PVY)、马铃薯卷叶病毒(PLRV)和马铃薯S病毒(PVS),开展了利用基因工程方法获得兼抗4种马铃薯病毒转基因马铃薯材料的研究。试验在前期获得含4种马铃薯病毒外壳蛋白基因片段的质粒pART27-XSYV-rh的基础上,通过根癌农杆菌(Agrobacterium tumefaciens)介导转化马铃薯(Solanum tuberosum)品种‘陇薯3号’,PCR扩增和PCR-Southern杂交证明,4价融合基因已整合到马铃薯基因组中。qRT-PCR分析表明,该融合基因在转基因植株中能正常表达。3株转基因植株的抗病性鉴定结果表明,2株对4种病毒同时具有抗性;1株对PLRV侵染表现阳性,对另外3种病毒同时具有抗性。 相似文献
4.
The incidence of Alfalfa mosaic virus (AMV), Potato leafroll virus (PLRV), and Potato virus Y (PVY) in potato crops derived from various types of seed potatoes was assessed visually and confirmed by direct tissue blot
immunoassay, over two winter growing seasons (1999/2000, 2000/2001) at three locations, Elnaiya, Elshehinab, and Shambat in
Khartoum State, the main potato growing region in Sudan. Virus infection was most prevalent in 2000/2001. In general, crops
grown directly from imported certified seed potatoes and from “improved seed”, produced in Sudan from imported basic seed,
showed the lowest levels of PLRV and PVY compared with crops grown from Sudanese farm saved seed. For AMV, however, only crops
grown directly from imported certified seed potatoes had low levels of AMV. Crop location also affected virus incidence, although
this varied with year. For AMV, levels were similar at all locations in 1999/2000, but were greatest at Elnaiya in 2000/2001.
For PVY, levels were greatest at Elnaiya in 1999/2000 and Shambat in 2000/2001. For PLRV, no symptoms were observed in 1999/2000
and virus levels were similar for all locations in 2000/2001. This study reports for the first time the occurrence of AMV
in potatoes grown in Sudan. 相似文献
5.
紫色甘薯的茎尖培养与脱毒 总被引：2，自引：0，他引：2
以紫色甘薯品系为试材,研究6-BA和NAA及基因型对紫色甘薯茎尖培养的影响,并采用硝酸纤维素膜—酶联免疫法(NCM-ELISA)和症状法对再生植株进行病毒学检测。结果表明,培养基中6-BA和NAA浓度和配比对紫色甘薯茎尖培养的愈伤组织、不定根和不定芽的诱导有明显影响。6-BA可促进不定芽的诱导,附加1mg/L6-BA的MS培养基为最佳诱芽培养基。基因型对诱芽率也有一定的影响,其中品系B3和B7的诱芽率可达50%。通过病毒学检测,获得12个紫色甘薯品系的脱毒苗,其平均脱毒率为94.7%。 相似文献
6.
The potato plantlets singly infected by PVA, PLRV, PVS, PVX and PVY and mix-infected by PVM, PVS and PVY were cultured on MS medium with different concentration of ribavirin. The effects of ribavirin on growth of the plantlets and efficiency of virus elimination were investigated. Results showed that the plant height and fresh weight obviously decreased with increase of ribavirin concentration from 0 mg/L to 150 mg/L, and most of the plantlets could not survive when the concentration reached 200 mg/L. According to the ELISA tests, ribavirin was more efficient for eradicating PVA, PVM, PVS and PVX than PVY and PLRV, and healthy plantlets could be obtained with high frequency (up to 100 %) by culturing with 75?~?150 mg/L ribavirin after 2?~?3 subcultures. Whereas, only 33?~?66 % PVY and PLRV infected plantlets were found to be virus-free after 3 subcultures with 75?~?150 mg/L ribavirin. The results of quantitative RT-PCR (qPCR) indicated that ribavirin could obviously reduce virus content in the plantlets. Except PLRV was detected positive after 3 subcultures with ribavirin, the healthy seedlings were obtained from infected stocks at the first or end of propagation and no viruses could be detected at the post-eradication stage. No apparent difference of genetic variation resulted from ribavirin treatment was found by SSR analysis between the control and the treated plantlets. All of these results above proved that ribavirin treatment in vitro was an effective method to eliminate viruses in the propagation of potato. 相似文献
7.
N. Ioannou 《Potato Research》1989,32(1):33-47
Summary The infection pressure of two viruses, potato leafroll (PLRV) and potato virus Y (PVY), both common in seed potatoes grown
in Cyprus, was determined in three experiments in 1982–83. Virus-free bait plants, of potato and four other species, were
exposed weekly to field infection during the growing season (March–June), and then returned to an aphid-free glasshouse for
symptom expression. Only tobacco plants produced clear symptoms enabling reliable assessment of PVY infection pressure. When
assessed with ELISA or by tuber indexing, the potato plants were efficient baits for both viruses whose infection period commenced
at emergence (mid March to early April) and ended within 6–7 weeks. The seasonal trend of aphid populations, determined with
Moericke traps or 100-leaf counts, correspond to that of virus spread. Correlation and regression analysis of aphid and virus
data implicated the alate form ofMyzus persicae as the principal vector of both viruses. 相似文献
8.
马铃薯茎尖脱毒效果影响因素的研究 总被引：6，自引：1，他引：5
以克新2号、克新16号和克新17号等8个马铃薯品种为试验材料,对马铃薯茎尖脱毒效果的影响因素进行了研究。结果表明:不同种类的植物生长调节剂对茎尖的生长有不同的影响,茎尖在⑤号培养基(MS+1.0mg·L-1KT+0.5mg·L-1IAA+0.5mg·L-1GA3)上可不产生愈伤组织,直接成苗,是最适宜的培养基;6～8℃低温预处理和37℃热空气处理对茎尖脱毒有利;茎尖取材来源对成苗率没有影响,对脱毒效果影响较大,但脱毒效果依品种而异;为避免造成损失,需在15个月内准备出用于更换的基础苗。 相似文献
9.
以马铃薯脱毒组培苗为试验材料,对N、P、K、Ca、Mg 5种元素的缺失、适量和加量而影响马铃薯脱毒组培苗生长状况进行试验研究。结果表明:各种元素的用量对组培苗的初生根时间影响较小,无明显差异;不同品系的脱毒组培苗对各种元素的需求量不同,总的来说,P、Ca、Mg的加量使用能够促进组培苗的生长;各种元素的缺失和加量对组培苗的黑尖、分枝等不良性状的影响以及对组培苗的有效叶片数、生根率、平均根长、平均株高等指标的影响存在明显的品系间差异。因此,建立马铃薯脱毒组培苗的快繁体系要根据繁育品系的不同,使用相应的优化培养基,这样既能减少马铃薯组培苗生长过程中的黑尖、分枝和苗弱等现象,又能促进生根和有效叶片的形成,保障和提高马铃薯脱毒组培苗的质量。 相似文献
10.
Potato virus Y (PVY) isolates from potato were divided into two major groups – I and II – on the basis of nucleotide sequence variability
at the 5′-UTR of the virus genome. In this paper, a consensus sequence for each group was compiled by examining PVY isolates
of potato. The consensus sequences of the two groups differed from each other by as much as 31% of nucleotide sequence divergence.
However, sequences of most individual members within each group (except for one, the 605 strain) were conserved and varied
by less than 1% in group II and up to 6% in group I. These results confirm that the two virus groups are markedly distinct
but that the 5′-UTR sequence of individual members within each group is highly conserved. These findings imply that the 5′-UTR
can be reliably applied for major grouping of isolates but that its use for discrimination among individual isolates within
each group is limited. Isolates of both group I and II included members of the O/N-NTN virus types. Classification of PVY
isolates in potato is discussed. 相似文献
11.
12.
不同处理方法对马铃薯茎尖成苗率的影响 总被引：2，自引：0，他引：2
试验以带马铃薯病毒的N88为材料,分别在40℃/25℃(4 h/20 h)的变温环境中,培养2、3、4周处理后茎尖剥离;以N88和D575试管苗为材料,在加入0,20,30 mg·L-1病毒唑的培养基中处理40 d后茎尖剥离。茎尖培养基为MS+0.05 mg·L-1 NAA+0.1 mg·L-1 6-BA+0.1 mg·L-1 GA3+4 g·L-1琼脂粉+30 g·L-1白砂糖的改良固体培养基,30 d后开始调查成苗率,每隔10 d调查1次,直到60 d,统计茎尖成苗率。结果表明:2周变温处理的茎尖成苗率和对照接近,但脱毒率有所提高;4周变温处理剥离茎尖成苗率最低,60 d成苗率仅为15%,脱毒率为100%;病毒唑浓度对不同材料的茎尖成苗率影响不同,其中D575经过20 mg·L-1病毒唑处理,茎尖培养60 d后成苗率最高达到83.3%,PVS脱除率为40%;材料N88在加入20 mg·L-1和30 mg·L-1浓度病毒唑的培养基中培养40 d后茎尖剥离,60 d成苗率分别为40.0%和41.7%,PVY、PLRV的脱毒率为100%。 相似文献
13.
Environmental factors influencing aphid transmission of potato virus Y and potato leafroll virus 总被引：1，自引：0，他引：1
Summary The effect of temperature, relative humidity (RH) and light on aphid transmission of potato virus Y (PVY) and potato leafroll
virus (PLRV) was studied using as vectorsMyzus persicae Sulz. andAphis gossypii Glov. Host susceptibility was enhanced by 48 h pre-inoculation exposure at 25°C and by 48 h post-inoculation exposure to
30°C. High RH (80%) in both pre- or postinoculation phases enhanced host susceptibility. Continuous fluorescent light (4000
lux) did not alter the rate of transmission of either virus. High RH (80–90%) and high temperature (25–30°C), when combined,
increased virus transmission by 30–35%. Transmission rates were reduced by nearly 50% if RH was maintained at 50% in either
of the two phases even if the temperature was 25 or 30°C. Both viruses were acquired by aphids earlier (13–20 days after inoculation)
when the source plants were incubated at 25 or 30°C. Most virus was transmitted from plants inoculated with PVY 13 to 16 days
and with PLRV 15 to 20 days previously. Transmission rates of PVY were enumerated from symptom expression on test plants and
by Enzyme Linked Immunosorbent Assay (ELISA) whereas those of PLRV were enumerated from symptom expression alone. 相似文献
14.
Potato is one of the most important crops worldwide. Genetic resources of potato (Solanum tuberosum L. ssp. tuberosum) and related cultivated species are conserved through storage of tubers, in vitro plants and in cryopreservation. Cryopreservation,
storage in or above liquid nitrogen, is the best option to maintain vegetatively propagated plants in the long term. The present
review gives comprehensive information about various cryopreservation techniques for potato published from 1977 until the
present. It discusses factors that affect the process and success of cryopreservation, such as donor culture conditions, preculture,
cooling, warming and post-culture treatments. Studies are presented that analyse the histological and ultrastructural changes
after different cryopreservation steps and the morphological pathways during regeneration of plants after rewarming. The maintenance
of genetic stability in potato after cryopreservation has also been demonstrated by various phenotypic and molecular methods.
The first thermal analyses on potato shoot tips are presented using differential scanning calorimetry to analyse the state
of water during cooling and warming. Biochemical analyses of different compounds, such as soluble sugars and proteins, have
been performed to understand and improve existing cryogenic methods. Potato is an example where successful virus elimination
has been obtained via cryopreservation of shoot tips (cryotherapy). There are already cryopreserved collections of potato
shoot tips in Germany, Peru, Czech Republic, South Korea and USA, but additional experiments on fundamental aspects of potato
cryopreservation will help to improve understanding of the different cryopreservation methods, start new collections in other
countries and also build up existing cryocollections of potato. 相似文献
15.
16.
Summary A greenhouse minituber production system involving low inputs of in vitro potato plantlets and propagation media is described,
in vitro plantlets of six potato cultivars were sectioned into nodal cuttings and separately planted into moist peat based
growing medium in shallow plastic trays. Cultivar differences were evident with respect to node viability, shoot regrowth
and minituber yields. Nodal viability for shoot regrowth varied between 80–100%. Maximum shoot heights were recorded with
whole in vitro plantlets (WIP) and the terminal Node-5 cluster. All cuttings produced minitubers. The terminal Node-5 cutting
and WIP produced significantly larger minitubers >3.0 g as compared to single node cuttings. Greater numbers of minitubers
were produced by the cvs Norchip, Red Pontiac and Conestoga as compared to cvs Eramosa. G8610-4PY and Shepody. Total numbers
of minitubers were 3 to 5 times higher from each in vitro plantlet that was sectioned into nodal cuttings as compared with
intact WIP: the yield ratios depended on cultivar. 相似文献
17.
In an experimental breeding scheme to improve late blight (Phytophthora infestans) and white potato cyst nematode (Globodera pallida) resistance of tetraploid potato over three generations of crossing and selection, 15 clones survived the final selection,
and these were derived from 15 great-grandparents. There was no direct selection for resistance to Potato virus Y (PVY), but 14 out of the 15 great-grandparents were resistant to PVY and three had extreme resistance. Thirteen of the 15
descendants had PVY resistance and one extreme resistance. This was within the range expected for a random (unselected) sample
from the genotypes of the great-grandparents. Hence, we found no evidence for any positive or negative association between
PVY resistance and the attributes selected. The conclusion is that laborious selection is not required in every generation
when many parents have PVY resistance, including some with more than one copy of a PVY resistance gene or resistance at more
than one locus. However, in the future, determining the major virus resistance genes present in potential parents in each
generation using diagnostic molecular markers would prevent susceptible × susceptible crosses being made and maximise the
number of resistant × resistant ones. 相似文献
18.
R. G. Novy A. Nasruddin D. W. Ragsdale E. B. Radcliffe 《American Journal of Potato Research》2002,79(1):9-18
Increasing prevalence of potato leafroll virus (PLRV) and potato virus Y (PVY) has been reported in seed and commercial potato production, resulting in the rejection of potatoes for certification and processing. Host plant resistance to PLRV and PVY and their primary vector, green peach aphid,Myzus persicae, could limit the spread of these viruses. Host plant resistance to PLRV, PVY, and green peach aphid has been identified in non-tuber-bearingSolanum etuberosum (PI 245939) and in its backcross 2 (BC2) progeny. Resistance to green peach aphid involved a reduction in fecundity and adult aphid size. In addition, one BC2 individual was identified as possessing a genetic factor that was detrimental to nymph survival. PVY resistance was identified in all five BC2 progenies evaluated in a field screening under intense virus pressure. PLRV resistance was identified in two of the five BC2 progeny. This resistance was stable in field and cage evaluations with large populations of viruliferous aphids. Based on the segregation of virus resistances in the BC2 , PVY and PLRV resistances appear to result from the action of independent genetic mechanisms that reduce the levels of primary and secondary virus infection. Two BC2 individuals, Etb 6-21-3 and Etb 6-21-5 were identified as having multiple resistances to PLRV, PVY, and green peach aphid derived fromS. etuberosum. This germplasm could prove useful to potato breeders in the development of virus-resistant cultivars. 相似文献
19.
Summary Uridine-H3 was incorporated into meristem tips of both healthy and PLRV-infected potato plants, of the cultivars, Majestic and Primura.
Autoradiograms of tips pretreated with actinomycin D showed that 13 of 14 and 11 of 14 respectively, were virus free in the
dome and first 4 leaf primordia, 1 and 3 were infected in the 4th leaf primordium, and 1 of Primura also in its 3rd primordium.
The highest plantlet yield from cultures in vitro of meristem tips that included 4 leaf primordia (7 to 9 plantlets per meristem),
was obtained by growing them in sequence on 3 sequential media, each based on Murashige and Skoog basic medium complemented
with various hormones. The 2nd medium, containing benzylaminopurine (0.5 mg/l) and gibberellic acid (0.5 mg/l), elicited callus
and the subsequent formation of adventitious buds. Of the plantlets of the cvs Vivaks, Primura and Majestic, 88, 91 and 100
%, respectively, were PLRV-free.
Propagation in vitro, by the single-node cutting technique and tuberlet production, were successful. 相似文献
20.
H. K. Were J. N. Kabira Z. M. Kinyua F. M. Olubayo J. K. Karinga J. Aura A. K. Lees G. H. Cowan L. Torrance 《Potato Research》2013,56(4):325-342
Potato plays an important role in food security in Kenya but yields are low (<10 t/ha), and this is partly attributed to the lack of healthy planting material. This study is the first wide-scale survey to determine the occurrence and distribution of common potato pests and diseases in Kenyan seed (certified and quality declared) and ware crops. Potato crops growing on 101 farms in 21 districts were examined. Approximately 36% of plants in farmers’ fields sampled both during the long rains (main potato-growing season) and short rains seasons displayed virus-like disease symptoms. Six viruses (potato leafroll virus (PLRV), Potato virus A (PVA), potato virus M (PVM), potato virus S (PVS), potato virus X (PVX), and potato virus Y (PVY)) were detected using double antibody sandwich enzyme-linked immunosorbent assay in potato samples. Sequencing of polymerase chain reaction products from PVY-infected plants revealed the presence of recombinant strains of PVY (NTN and Wilga). Four aphid species, Macrosiphum euphorbiae, Aphis gossypii, Myzus persicae, and Aphis fabae, colonized potato in all districts, occurring in greater numbers west of the Great Rift Valley than to the east. There was a positive correlation between virus incidence and aphid numbers in the long rains (main) potato-growing season. PLRV, PVM, PVS, PVX, and PVY were detected in solanaceous weeds. Ralstonia solanacearum was detected in soils from 13 farms in 8 of the 18 districts surveyed. Approximately 38% of soil samples were infested with Meloidogyne spp. Phytophthora infestans isolates belonging to the US 1 and 2_A1 genotypes were identified. Although many economically important diseases are present in Kenya, the lower aphid incidence in districts east of the Great Rift Valley may indicate that these districts are more suitable for seed potato production. 相似文献