PSTV与PVY的互作及其对马铃薯产量影响

本文分别在温室和田间条件下利用人工接种的方法研究了马铃薯纺锤块茎类病毒（ＰＳＴＶ）和ＰＶＹ在克新４号及Ｋｅｎｎｅｂｅｃ两品种上的相互作用．试验结果表明：在温室条件下ＰＳＴＶ和ＰＶＹ的复合接种均能在二品种上不同程度地产生条型坏死症状，而在田间未出现明显症状．ＰＳＴＶ与ＰＶＹ混合接种或ＰＳＴＶ接种一周后再接ＰＶＹ会促进ＰＶＹ增殖，植株内ＰＶＹ的浓度显著高于单独接种ＰＶＹ，但ＰＶＹ侵染后再接ＰＳＴＶ对ＰＶＹ的浓度无显著影响．ＰＳＴＶ和ＰＶＹ的工作对ＰＳＴＶ的浓度无显著影响．ＰＳＴＶ与ＰＶＹ复合侵染的三个接种类型都明显地降低平均单株产量和商品薯率，产量损失最大的处理为ＰＳＴＶ与ＰＶＹ混合接种，可导致克新４号减产４５．４％。     

应用NASH方法检测马铃薯类病毒(PSTVd)

1　前　言马铃薯纺锤块茎类病毒 (ＰｏｔａｔｏＳｐｉｎｄｌｅＴｕｂｅｒＶｉｒｏｉｄ ,简称ＰＳＴＶｄ)是引起马铃薯纺锤块茎病的原因 ,该病害发现于 192 2年 ,但直到 196 7年及以后的研究中才发现该病由一种叫类病毒的病原引起的。分布十分广泛 ,是危害我国北方马铃薯生产的重要病害之一 ,严重影响马铃薯的质量和产量 ,一般弱系可造成减产 2 0 %～ 35 % ,强系可达 6 0 %。对其检测有指标植物方法 (利用指示植物鲁格斯番茄Ｒｕｔｇｅｒｓ ,新莨蓿等 ) ,在我国 ,目前较常用的检测方法为聚丙烯酰胺凝胶电泳法 ,而随着分子生物学的发展 ,杂交…     

培育健壮马铃薯试管苗试验

我们在马铃薯茎尖脱毒和试管苗繁殖过程中发现，马铃薯试管苗生长细弱、茎叶嫩黄与带有高浓度的马铃薯纺锤块茎类病毒（PSTV）及几种主要马铃薯病毒有关。用双向聚丙烯酰胺凝胶电泳法和酶联夹心法检测筛选出无病毒的试管苗作为试材，排除类病毒和病毒侵染的干扰，以提高培养基中营养元素含量作为培养健壮试管苗的措施，试验结果表明，2倍MS液体培养处理的试管苗生长茁壮、浓绿，加速了干物质累积、使继代培养周期从3～4周缩短为2～3周。     

培育健壮马铃薯试管苗试验

我们在马铃薯茎尖脱毒和试管苗繁殖过程中发现，马铃薯试管苗生长细弱、茎叶嫩黄与带有高浓度的马铃薯纺锤块茎类病毒（ＰＳＴＶ）及几种主要马铃薯病毒有关。用双向聚丙烯酰胺凝胶电泳法和酶联夹心法检测筛选出无病毒的试管苗作为试材，排除类病毒和病毒侵染的干扰，以提高培养基中营养元素含量作为培养健壮试管苗的措施，试验结果表明，２倍ＭＳ液体培养处理的试管苗生长茁壮、浓绿，加速了干物质累积，使继代培养周期从３～４周缩短为２～３周。     

马铃薯生产中马铃薯纺锤块茎类病毒的综合防治

马铃薯纺锤块茎类病毒是影响中国马铃薯生产重要的检疫性病害。该病害具有广泛的自然寄主,可侵染马铃薯、番茄和辣椒等重要作物。该病害可通过植物学种子、花粉或卵细胞、无性繁殖以及机械等方式传播。马铃薯纺锤块茎类病毒可造成马铃薯产量降低,块茎畸形、变小,商品性和商品薯率下降。该病害主要可通过培育抗病品种、使用脱毒种薯、加强种质资源和种薯卫生管理、加强检验检疫以及实行马铃薯种薯质量认证制度等措施进行综合防治。     

不同品种（系）马铃薯休眠块茎顶端和茎端芽眼组织马铃薯Y病毒含量分析

马铃薯Y病毒（Potato virus Y,PVY）是马铃薯生产中较常见的病毒之一,马铃薯病毒的检测是生产中保障马铃薯品质和产量的重要基础,收获后批量检测是进行种薯质量评价的主要依据之一。研究针对‘56-2’‘Ivory Russet’‘Clearwater Russet‘’龙薯14号‘’龙薯3号‘’龙薯15号‘’龙育201401-70‘’龙薯7号‘’克新23号’和‘克新28号’马铃薯品种（系）休眠块茎样品,采用TRIzol法提取马铃薯休眠块茎顶端和茎端芽眼组织的总RNA,经qRT-PCR和RT-PCR检测分析,比较块茎的顶端与茎端芽眼组织PVY含量分布的情况。结果表明,上述供试材料的休眠块茎茎端芽眼组织PVY浓度均高于顶端芽眼组织,能够精准的检测到病毒的存在;对‘Ivory Russet’品种的休眠块茎选取100个带病样品通过TRIzol法提取顶端与茎端的总RNA,采用4合1的方法合样后进行PVY RT-PCR检测,所有处理的顶端芽眼组织检测条带亮度低于茎端芽眼组织。可见,马铃薯休眠块茎茎端芽眼组织取样能够更加精准的检测出PVY。研究结果为马铃薯种薯收获后PVY的检测及合理取样提供了...     

R-PAGE胶板的干燥方法

１　前　言马铃薯纺缍块茎类病毒（ｐｏｔａｔｏｓｐｉｎｄｌｅｔｕｂｅｒｖｉｒｏｉｄ）是一种严重危害马铃薯生产的类病毒，降低产量２０％～３０％。防治的主要措施是应用无病毒种薯。由于目前还没有脱掉类病毒的有效措施，只能从未被饱和侵染的群体中鉴定筛选出未被侵染的个体，再脱掉其它病毒，作为核心材料。为了能够筛选出更多的无马铃薯纺缍块茎类病毒（ＰＳＴＶ）的个体，确保种薯生产和无毒化育种的质量，必须加强马铃薯纺缍块茎类病毒的检测工作。目前我们采用的检测马铃薯纺缍块茎类病毒的方法是往复—聚丙烯酰胺凝胶电泳法（Ｒ…     

不同纬度引种对马铃薯产量和淀粉品质的影响

影响马铃薯引种成功与否的因素有很多,如纬度、海拔、气候条件和品种生育期等。本试验选取当地主栽马铃薯品种‘龙引薯1号’和‘克新13号’,分别来源于不同纬度地区呼玛县、嫩江县和林甸县的种薯,引种到林甸县,测定生长期间的株高和茎粗、收获后的块茎产量和大中薯率,淀粉含量,提取淀粉产量并测定淀粉黏度和糊化温度,以分析比较不同纬度引种对马铃薯植物学性状、产量及品质的影响。结果表明,两个品种均表现为来源于高纬度地区的种薯长出的马铃薯植株高大、茎秆粗壮,产量高、大中薯率高,淀粉产量也高,但淀粉提取率、淀粉黏度及糊化温度变化规律不明显。     

部分发展中国家马铃薯纺锤块茎类病毒病情况调查

马铃薯类病毒病是威胁世界马铃薯生产的主要病害之一,更影响着发展中国家马铃薯产业的发展。2008~2010年,黑龙江省农业科学院植物脱毒苗木研究所连续3年举办了马铃薯病害检测技术国际培训班,对部分发展中国家从事马铃薯工作的人员进行培训,在培训过程中进行了马铃薯类病毒方面的问卷调查,调查结果显示:这些发展中国家对马铃薯纺锤块茎类病毒比较了解,发病较重的国家不多,但总体检测水平不高,甚至不进行马铃薯类病毒的检测,多数国家不重视马铃薯纺锤块茎类病毒病的防治工作。因此,马铃薯纺锤块茎类病毒病是发展中国家的隐患。     

反复冷冻后块茎组织中马铃薯纺锤块茎类病毒的损失

许多国家均认为马铃薯纺锤块茎类病毒(PSTV)是一个危险的病原。因此,出口到一些国家的种薯需要进行PSTV检验,这种检验通常是以实验室检测为基础的,从每批种薯中随机抽取300～1000个薯块,检测PSTV和细菌环腐病。对于这样多的待测薯样,通常是从待测薯块上取下一小部分,     

Comparative detection of mild strains of potato spindle tuber viroid from the dormant potato tubers by Return-polyacrylamiee gel electrophoresis and nucleic acid hybridization

Summary Using a modified procedure of Return-polyacrylamide gel electrophoresis (R-PAGE) mild strains of potato spindle tuber viroid (M-PSTV) were detected reliably from dormant tubers. The sensitivity of R-PAGE detection of M-PSTV was equivalent to that of nucleic acid hybridization. Both methods detected M-PSTV when infected tissue was mixed with healthy tissue in a ratio of 1 to 100. When extracted nucleic acid was diluted with buffer, R-PAGE detected PSTV at a dilution of 1:256 and nucleic hybridization only up to 1:64 PSTV was readily detected from 18 potato cultivars. In addition, mild, ‘intermediate’ and severe strains were separated by R-PAGE, on the basis of their mobility on the electrophorogram.     

Detection of potato spindle tuber viroid by nucleic acid spot hybridization: Evaluation with tuber sprouts and true potato seed

A diagnostic test for the potato spindle tuber viroid (PSTV) that is based on hybridization of highly radioactive, recombinant DNA complementary to PSTV with PSTV bound to a nitrocellulose membrane and autoradiographic detection of the resulting DNA-RNA hybrids has been evaluated with tubers from 20 potato clones maintained at the International Potato Center (CIP) and with true seed obtained from healthy or PSTV-infected potato plants. The nucleic acid spot hybridization test detected PSTV not only in tuber sprouts from 10 clones that had previously tested positive in polyacrylamide gel electrophoretic (PAGE) analysis, but also in tuber sprouts from 9 clones that had previously tested negative in PAGE analysis. Further tests confirmed the presence of PSTV in these clones. The spot hybridization test detected PSTV in mixtures of seed extracts equivalent to as few as one seed from a PSTV-infected plant and 80 seeds from healthy plants. The spot hybridization test was shown to be more sensitive and reliable than PAGE analysis; it is suitable for the testing of large numbers of samples with a minimum expenditure of labor and materials.     

New disease symptoms observed on field-grown potato plants with potato spindle tuber viroid and potato virus Y infections

Summary Potato spindle tuber viroid (PSTV) and potato virus Y (PVY) were isolated from plants of cultivar Kennebec with severe necrotic symptoms in the field. In the greenhouse, severe necrotic symptoms were reproduced only when potato plants were infected either simultaneously with PSTV+PVY, or with PSTV prior to PVY infection. Thirteen additional potato cultivars were tested in the greenhouse for this synergistic reaction and eight developed necrotic responses similar to cv. Kennebec. PVY concentration was significantly higher in doubly infected plants, compared with those infected with PVY alone.     

Ineractions between a mild and a severe strain of potato spindle tuber viroid in doubly infected potato plants

Potato (Solarium tuberosum) plants co-infected with a mild and a severe strain of potato spindle tuber viroid (PSTVd) were analyzed by return-polyacrylamide gel electrophoresis for the presence of both strains in vegetative and reproductive plant parts. Both strains were detected in the anthers, flowers, inflorescences, leaves, ovaries, ovules, petals, pistils, roots, sepals, stolons and tubers. Only mild strain was detected from pollen in the cultivars tested. True potato seed (TPS) were not doubly infected when they were obtained from co-infected maternal parent plants pollinated with pollen from healthy plants. Also, when maternal plants infected with severe strain were pollinated with pollen from healthy plants or from those infected with the mild strain, TPS were not doubly infected. A small number of TPS with double infection was obtained when co-infected or mild-strain-infected plants were pollinated with pollen containing the severe strain (singly or doubly infected). The number of TPS containing mild strain predominated in a ratio of 7:1 over TPS containing the severe strain. This study indicates that segregation of strains from doubly-infected plants probably takes place during pollen formation and persists through seed development.     

Evaluation of six insect pests for transmission of potato spindle tuber viroid

Six insect pests of potato, green peach aphid,Myzus persicae (Sulzer); potato leafhopper,Empoasca fabae (Harris); Colorado potato beetle,Leptinotarsa decemlineata (Say); tarnished plant bug,Lygus lineolaris (Palisot de Beauvois); red-legged grasshopper,Melanoplus femurrubrum (DeGeer); and southern army worm,Prodenia eridania (Cramer), were tested for their ability to transmit potato spindle tuber viroid (PSTV). PSTV infection was determined by a gel electrophoretic assay made 6 weeks after transfer of insects from infected to non-infected plants of the cultivar, ‘Katahdin.’ Plants grown from tubers of the original non-infected hosts were also assayed. PSTV was detected in only 2 of 183 test plants; these 2 plants had been infested with the tarnished plant bug. These results suggest that the insect species studied are relatively insignificant as vectors of PSTV.     

Loss of potato spindle tuber viroid from tuber tissues after repeated freezing

Potato spindle tuber viroid (PSTV) infected foliage and tuber tissues when subjected to repeated freezing and thawing under controlled conditions (?18 to ? 20 C and 5 C), lost PSTV more rapidly in tuber tissues than in foliage. Tubers of 17 potato cultivars showed similar tendencies, but with some differences. In 7 of the 17 cultivars, PSTV was significantly reduced after one freeze-thaw treatment, and in 6 of the 17 cultivars, infectious PSTV was still recoverable after exposure to 4 cycles of freeze-thaw. Exposure of PSTV-infected tubers to outdoor temperature fluctuations (13.1 C to ?15.1 C) resulted in complete loss of PSTV from the tubers.     

Detection,distribution and long-term persistence of potato spindle tuber viroid in true potato seed from heilongjiang,China

Return-polyacrylamide gel electrophoresis (R-PAGE) and tomato bioassay followed by R-PAGE were compared for the detection of potato spindle tuber viroid (PSTVd) from individual true potato seed (TPS). Both methods detected PSTVd from single TPS. TPS extract formulated as sap or nucleic acids in two different buffers did not affect the percentage of viroid detection on tomato plants. There was some evidence of viroid inhibitor in TPS extracts but not in nucleic acid extracts of TPS. Because R-PAGE is more rapid than the tomato bioassay followed by R-PAGE, the former was used to determine the extent of PSTVd in TPS of China’s Keshan Potato Research Institute breeding material. Over 1700 individual TPS were tested. Twenty-four of the 46 seedlots tested (inbred and outcrossed) contained PSTVd. The viroid was detected in 70% of lots from inbred lines compared to 38% of lots from outcrosses. TPS (20 lots) stored in paper bags at room temperature as far back as 1965 were also tested, and PSTVd was detected in TPS stored for 21 years.     

Purple top roll disease of potato

A graft transmissible disease of potato termed purple top roll is prevalent in several potato varieties grown in the hilly tracts of India. The common symptoms are rolling of the basal parts of younger leaves, sometimes accompanied by pink or purple pigmentation. The plants are stunted with profuse growth of axillary branches and aerial tubers. The losses in yield varied from 40 to 70%. Five to 20% of the tubers from diseased plants developed hairy sprouts. Tuber transmission of the pathogen varied from 6 to 33%, depending on the variety.Datura, tomato and white clover are among susceptible plants.N. rustica was tolerant or a symptomless carrier. Sunnhemp, Petunia andSesamum were immune to the pathogen, which was transmitted by the leaf hopper,O. albicinctus Distant but not byCuscuta spp. or by sap inoculation. Application of Chloramphenicol, Tetracycline and Antiamoebin resulted in remission of symptoms and increased tuber yields, indicating possible association of a mycoplasma-like organism.     

Studies onFusarium wilt of potatoes. 2. Leaf,sprout and tuber infection in artificial inoculations

Summary In artificial inoculations it was found thatFusarium oxysporum f.sp.tuberosi is able to penetrate through potato leaves. The fungus was isolated from the stem of infected plants from all inoculated cultivars. Tubers were completely destroyed if they were inoculated before sprouting. Not any destruction was observed if inoculation was made before young sprouts are longer than 1–3 cm; plants from these sprouts were infected. Lesioned tubers were more sensitive than unlesioned ones, and tuber rot and sprout damage were increased significantly.