棉蚜获毒后禁食对其保持并传播黄瓜花叶病毒的影响

采用棉蚜Aphis gossypii 甜瓜Cumumis melo 黄瓜花叶病毒（cucumber mosaic virus, CMV）体系,研究棉蚜获毒后在空气中禁食对其保持并传播黄瓜花叶病毒的影响。结果表明获毒后的禁食时间与棉蚜传毒效率呈负相关。运用EPG (electrical penetration graph)及其即时显示、即时中断技术研究分析棉蚜禁食后的早期传毒行为细节。结果显示：禁食处理没有显著影响电势落差（potential drop,pd）数目及穿刺过程中出现的第一个pd波形前穿刺时间这两个重要指标,但禁食处理能引起pd波的两个亚波形pdⅡ-1和pdⅡ-2持续时间的显著减短。进一步分析未禁食棉蚜传毒作用与pd亚波形的关系,显示传毒可能与pdⅡ-2的持续时间相关（P＝0.06）。因此,pdⅡ-2的持续时间可能是与棉蚜传毒相关的一个行为指标。该研究还建立了新的高效而稳定的获毒方法---5pd获毒法,与传统的5min获毒法相比,获毒效率显著提高。     

石榴园棉蚜及其天敌之间的关系

采用灰色系统分析方法,分析了对主要害虫棉蚜及其天敌日捕食总量之间关联度,结果表明,棉蚜群落中与其关联度较高的天敌种类依次为肖蛸蛛(0．8607)、中华草蛉(0．8058)、八斑球腹蛛(0．7989)、异色瓢虫(0．7881)和大草蛉(0．7758),与理想天敌日捕食总量间关联度较高的天敌种类为草间小黑蛛(0.8975)、智利小绥螨(0．8132)、龟纹瓢虫(0.7806)和大草蛉(0．7669);与天敌数量间关联度较高的天敌依次为草间小黑蛛(0．8482)、中华草蛉(0．7533)、肖蛸蛛(0．7532)、八斑球腹蛛(0．7411)和大草蛉(0．7116),与理想天敌数量间关联度较高的为草间小黑蛛(0．8461)、智利小绥螨(0．7325)、龟纹瓢虫(0．6983)、大草蛉(0．6815)和中华草蛉(0．6757)．棉蚜垂直生态位和水平生态位的重迭值均>0．9567的天敌有草间小黑蛛、大草蛉和肖蛸蛛;时间生态位重迭值>0．4020的天敌有草间小黑蛛、大草蛉和肖蛸蛛,表明草间小黑蛛、大草蛉、肖蛸蛛等为棉蚜主要天敌优势种。     

Resistance of transgenic tomato to cucumber mosaic cucumovirus under field conditions

Since the summer of 1993, transgenic tomato plants expressing the coat protein (CP) genes of cucumber mosaic cucumovirus have been tested under field conditions to assess the level of resistance and agronomic performance. Trials were performed in different areas in Italy and the target virus in the majority of tests was spread naturally by the indigenous aphid populations. Twenty-three homozygous lines of variety UC82B, transformed to contain four different CP genes of CMV, were evaluated. The lines were preselected for CP expression, single gene copy, and virus resistance in growth chamber experiments. In general, CMV resistance was confirmed under field conditions though resistance in the field was less effective than what was observed in growth chamber experiments. The resistance observed in multi-year and multi-location experiments is of commercial value for several of the most resistant lines. Engineered resistance upon transfer to Italian varieties by breeding or direct transformation will be used in tomato production in Italy or elsewhere. This revised version was published online in June 2006 with corrections to the Cover Date.     

黄瓜花叶病毒基因组不同部分及卫星RNA介导的对植株的保护及其作用机理

根据病原物介导的对自身抗性的理论,大量开展了将CMV基因组的单个或多个片断转入植物体内的研究,从而使该植株能够抵抗或延迟受CMV的侵染,CP,RP,MP基因是CMV基因组的重要组成部分,用来转化植株取得了不同程度的抗性效果,另外有些CMV株中存在着起致弱作用的卫星RNA,直接对植株接种含卫星RNA的CMV弱毒或用卫星RNA的cDNA转化植株都会减轻CMV强毒对该植株的侵害,CMV基因组不同组分进入植物体内后,它们对植株产生保护作用的机理不同,文中分别加以阐述。     

Evolutionary relationship of alfalfa mosaic virus with cucumber mosaic virus and brome mosaic virus

The amino acid sequences of the non-structural protein (molecular weight 35,000; 3a protein) from three plant viruses — cucumber mosaic, brome mosaic and alfalfa mosaic have been systematically compared using the partial genomic sequences for these three viruses already available. The 3a protein of cucumber mosaic virus has an amino acid sequence homology of 33.7% with the corresponding protein of brome mosaic virus. A similar protein from alfalfa mosaic virus has a homology of 18.2% and 14.2% with the protein from brome mosaic virus and cucumber mosaic virus, respectively. These results suggest that the three plant viruses are evolutionarily related, although, the evolutionary distance between alfalfa mosaic virus and cucumber mosaic virus or brome mosaic virus is much larger than the corresponding distance between the latter two viruses.     

棉花型和瓜型棉蚜产生有性世代能力的分化

为了明确寄主专化性是否会影响棉蚜的繁殖策略或生活史对策,采用低温和短光照组合（18℃,L∶D=8∶16）分别对棉花型和瓜型棉蚜进行有性世代的诱导,比较两寄主专化型棉蚜产生有翅蚜、雄性母、雌性母及无翅孤雌蚜的能力。结果表明：两寄主专化型棉蚜在产生有性世代能力上存在显著差异,表现为棉花型棉蚜在诱导后代中产生有翅蚜、雄性母和雌性母的比率显著高于瓜型棉蚜,并且雌、雄性母产生的时间明显早于瓜型棉蚜。瓜型棉蚜在诱导条件下产生无翅孤雌蚜的比率显著高于棉花型棉蚜,并且发现有不产生有性世代的营专性孤雌生殖个体,而在棉花型棉蚜中没有发现。产生性母蚜的棉花型和瓜型棉蚜均可同时产生孤雌胎生后代。寄主型与诱导时间长短对棉蚜有性世代的产生存在交互作用。采集于田间棉花和黄瓜上的棉蚜,也表现为棉花上的易产生有性世代,在诱导的第2代中产生性母蚜的比率显著高于黄瓜上的棉蚜,并且性母蚜比率在诱导的第2代与第3代间无显著差异; 但黄瓜上的棉蚜性母蚜产生比率第3代显著高于第2代。由此推测,棉蚜寄主专化性的形成与生活史特性的分化有关。     

棉蚜抗吡虫啉品系和敏感品系主要解毒酶活性比较

通过生物测定和生物化学方法比较了棉蚜 Aphis gossypii 对吡虫啉抗性（约为7倍）和敏感品系几种主要解毒酶的活性。结果表明：氧化胡椒基丁醚对两个品系均无明显增效作用。抗性品系中羧酸酯酶和谷胱甘肽S-转移酶的比活力均明显高于敏感品系,抗性品系中羧酸酯酶的K_m值也显著高于敏感品系,说明抗性品系羧酸酯酶与底物的亲和力明显高于敏感品系。上述结果证明羧酸酯酶和谷胱甘肽S-转移酶活力增强在棉蚜对吡虫啉的抗性中起重要作用。     

有机农业棉田中棉蚜及其捕食性天敌的种群动态

采用定期目测棉叶上的棉蚜数量,用扫网法同步估测天敌种群数量,对有机农业棉田和常规棉田(对照)中棉蚜及其捕食性天敌的种群动态进行了比较。结果表明,有机棉田和常规棉田中棉蚜及其主要捕食性天敌种群动态差异显著。在2005年6~8月,有机棉田中棉蚜数量是常规棉田的4.17倍(P<0.05)。瓢虫成虫和幼虫的数量分别是常规棉田的3.22倍(P<0.01)和3.70倍(P>0.05);草蛉成虫和幼虫的数量分别是常规棉田的1.33倍(P>0.05)和3.00倍(P<0.05);食蚜蝇的数量是常规棉田的2.89倍(P>0.05),蜘蛛的数量是常规棉田的3.47倍(P<0.05)。因此,建议在有机农业棉花生产中,采用一些植物源杀虫剂辅助控制棉蚜。     

Extreme resistance to cucumber mosaic virus (CMV) in transgenic tomato expressing one or two viral coat proteins

For the production of broad commercial resistance to cucumber mosaic virus (CMV) infection, tomato plants were transformed with a combination of two coat protein (CP) genes, representing both subgroups of CMV. The CP genes were cloned from the CMV-D strain and Italian CMV isolates (CMV-22 of subgroup I and CMV-PG of subgroup II) which have been shown to produce severe disease symptoms. Four plant transformation vectors were constructed: pMON18774 and pMON18775 (CMV-D CP), pMON18831 (CMV-PG CP) and pMON18833 (CMV-22 CP and CMV-PG CP). Transformed R0 plants were produced and lines were selected based on the combination of three traits: CMV CP expression at the R0 stage, resistance to CMV (subgroup I and/or II) infection in growth chamber tests in R1 expressing plants, and single transgene copy, based on R1 segregation. The results indicate that all four vector constructs generated plants with extremely high resistant to CMV infection. The single and double gene vector construct produced plants with broad resistance against strains of CMV from both subgroups I and II at high frequency. The engineered resistance is of practical value and will be applied for major Italian tomato varieties. This revised version was published online in June 2006 with corrections to the Cover Date.     

抗吡虫啉棉蚜种群对吡蚜酮等药剂的交互抗性及施药对其生物学特性的影响

为了明确吡蚜酮对抗吡虫啉棉蚜Aphis gossypii种群的防治效果,提出抗吡虫啉棉蚜的治理策略,利用抗吡虫啉棉蚜种群(RF₂₇)、敏感种群(SS)和夏津田间种群(XJ),分别采用浸渍法、微量点滴法、生化测定法和系统观察法研究了棉蚜无翅成蚜对吡蚜酮等药剂的交互抗性及施药对其生物学特性的影响。结果表明: 吡蚜酮对RF₂₇,XJ和SS的LD₅₀分别为1.213×10^-5,8.506×10^-5和5.140×10^-5 μg/头,RF₂₇对吡蚜酮表现出明显的负交互抗性现象。RF₂₇对啶虫脒、烯啶虫胺和噻嗪酮分别产生2.35,2.98和1.71倍的抗性。RF₂₇的羧酸酯酶和谷胱甘肽-S-转移酶的比活力较SS分别高2.73和1.57倍,说明羧酸酯酶和谷胱甘肽-S-转移酶比活力的提高是引起棉蚜对吡虫啉产生抗性的重要原因之一。吡蚜酮分别处理RF₂₇,XJ和SS,其羧酸酯酶和谷胱甘肽-S-转移酶均受到显著抑制。吡蚜酮以低剂量处理RF₂₇成蚜,对其生长发育有显著的不利影响,表现为若蚜存活率降低(64.60%),净生殖率降低（10.39）,内禀增长率和周限增长率显著降低(分别为0.21和1.23),世代历期延长（10.87 d）,相对适合度较小（仅为0.70）。这些结果表明吡蚜酮在棉蚜防治中具有很高的应用价值。     

Anti-sense RNAs of cucumber mosaic virus in transgenic plants assessed for control of the virus

Three synthetic genes for the production of anti-sense RNA to different regions of the cucumber mosaic virus (CMV) genome were constructed using virus-derived double-stranded cDNA coupled to a promoter sequence from cauliflower mosaic virus. The genes were used to transform tobacco plants by a Ti plasmid vector. Transgenic plants obtained with the three constructs produced anti-sense RNA at different levels. Plants expressing each of the three anti-sense RNAs were inoculated with CMV and their sensitivity to the virus infection was compared with the non-transformed plants. Only one plant line which expressed relatively low levels of one of the anti-sense RNAs showed resistance to CMV but other plants expressing the same or the other two antisense RNAs had similar sensitivity to CMV infection as the non-transformed plants.     

Rapid production and field testing of homozygous transgenic tobacco lines with virus resistance conferred by expression of satellite RNA and coat protein of cucumber mosaic virus

A procedure for the fast production of homozygotic transgenic plants was developed. Leaf discs of haploid tobacco plants from anther cultures were transformed with a chimaeric vector containing coat protein (CP) and satellite RNA (Sat-RNA) genes from cucumber mosaic virus (CMV). One-hundred-and-twelve Kanamycin-resistant transformed haploid plants were subjected to selection based on the expression of both CP and Sat-RNA. Eighty-nine transgenic plants expressing both genes were selected and tested for their resistance to CMV by inoculation with high concentration of CMV (200 g ml^–1). Only five plants showed no symptoms of viral infection 30 days after inoculation. These plants were then diploidized by colchicine treatment. Three homozygous diploid lines with high levels of resistance to CMV were obtained after only one generation. The three transgenic lines were further tested under field conditions. The results showed that the progenies of these transgenic lines were homozygous and were highly resistant to CMV under natural field infection and manual inoculation conditions.     

抗吡虫啉棉蚜种群对啶虫脒和噻虫胺的交互抗性及相关酶学机理

【目的】通过对乙酰胆碱受体β1亚基突变后的抗吡虫啉棉蚜Aphis gossypii (Glover)种群的继续筛选, 明确该种群的抗性发展规律以及对其他新烟碱类杀虫剂啶虫脒和噻虫胺的交互抗性及相关酶学机理。【方法】采用浸渍法连续对抗吡虫啉棉蚜进行室内筛选、 测定噻虫胺和啶虫脒对抗吡虫啉棉蚜种群的毒力; 选择LC₂₀剂量吡虫啉、 啶虫脒和噻虫胺处理抗性棉蚜, 采用生化分析法测定其体内羧酸酯酶、 谷胱甘肽-S-转移酶和乙酰胆碱酯酶的活性变化, 并观察其生物学特性的变化。【结果】本研究对抗性棉蚜突变种群用吡虫啉继续筛选至75代, 抗性倍数达到72.6倍, RF75停止用药筛选12代（RF₇₅₊₁₂）, 抗性仍达72.0倍。且RF₇₅₊₁₂对噻虫胺和啶虫脒的交互抗性可分别达11.9倍和20.1倍。噻虫胺对抗吡虫啉棉蚜的蜜露分泌和体重的抑制作用均大于吡虫啉和啶虫脒。噻虫胺对RF₇₅₊₁₂的羧酸酯酶、 谷胱甘肽-S-转移酶和乙酰胆碱酯酶均具有明显的抑制作用, 而啶虫脒的抑制作用较小。【结论】结果表明乙酰胆碱受体基因突变棉蚜种群对吡虫啉的抗性水平不仅升高, 且停止用药后其抗性可稳定遗传; 第二代新烟碱类的噻虫胺在抗吡虫啉棉蚜靶标突变种群的治理中具有较大的应用价值。     

Induction of resistance in tomato against cucumber mosaic cucumovirus by plant growth-promoting rhizobacteria

Studies were done to evaluate specific strains of plant growth promoting rhizobacteria (PGPR) for induced resistance against cucumber mosaic cucumovirus(CMV) in tomato. In greenhouse experiments where plants were challenged by mechanical inoculation of CMV, the percentage of symptomatic plants in the most effective PGPR treatments ranged from 32 to 58%,compared with 88 to 98% in the nonbacterized, challenged disease control treatment. Field experiments were conducted in 1996 and 1997 to evaluate 4 PGPR strain treatments based on superior performance in the greenhouse studies. In the 1996field experiment, tomato plants treated with 3 PGPR strains exhibited a significantly lower incidence of CMV infection and significantly higher yields, compared with nonbacterized, CMV-challenged controls. In 1997, the overall percentages of plants infected with CMV in the control and PGPR treatments was higher than in 1996. CMV symptom development was significantly reduced on PGPR-treated plants in 1997compared with the control, but the percentage of infected plants and tomato yields were not significantly different among treatments. These results suggest that PGPR-mediated induced resistance against CMV infection following mechanical inoculation onto tomato can be maintained under field conditions. This revised version was published online in July 2006 with corrections to the Cover Date.     

The potential use of a viral coat protein gene as a transgene screening marker and multiple virus resistance of pepper plants coexpressing coat proteins of cucumber mosaic virus and tomato mosaic virus

Transgenic pepper plants coexpressing coat proteins (CPs) of cucumber mosaic virus (CMV-Kor) and tomato mosaic virus (ToMV) were produced by Agrobacterium-mediated transformation. To facilitate selection for positive transformants in transgenic peppers carrying an L gene, we developed a simple and effective screening procedure using hypersensitive response upon ToMV challenge inoculation. In this procedure, positive transformants could be clearly differentiated from the nontransformed plants. Transgenic pepper plants expressing the CP genes of both viruses were tested for resistance against CMV-Kor and pepper mild mottle virus (PMMV). In most transgenic plants, viral propagation was substantially retarded when compared to the nontransgenic plants. These experiments demonstrate that our transgenic pepper plants might be a useful marker system for the transgene screening and useful for classical breeding programs of developing virus resistant hot pepper plants.     

棉花型和瓜型棉蚜形态和生态适应力的分化

棉蚜由于长期在同种或同类植物上生活,形成了多种寄主专化型。本研究以室内长期用单一寄主培养的棉花型和瓜型棉蚜为材料,采用形态测定、生命表技术、对寄主的选择性试验,以及接触刺激等方法,研究了两寄主型棉蚜的形态分化和生态适应力的差异。结果表明,瓜型棉蚜的一些形态特征值（如体长）极显著地大于棉花型个体,但是头宽、口针和各腿节的长度与体长的比值却极显著的小于棉花型个体。采用7个形态指标与体长的比值,可将瓜型和棉花型棉蚜区分开来。在24℃和26℃下瓜型蚜的净增殖率（R₀）极显著地高于棉花型蚜。27℃和32℃显著缩短了瓜型蚜的平均世代长度,而对棉花型蚜无影响。棉花型蚜对接触刺激的反应比瓜型蚜敏感。两寄主型蚜对寄主的选择能力表现为瓜型蚜显著强于棉花型蚜。因此,两寄主型棉蚜存在明显的形态和生态适应力的分化。     

棉蚜对吡虫啉的抗性选育和现实遗传力分析

【目的】为了评估棉蚜Aphis gossypii Glover对吡虫啉的抗性风险,在室内进行了棉蚜对吡虫啉(imidacloprid)的抗性选育和抗性现实遗传力分析。【方法】采用单头反选育法和群体汰选法,分别得到了棉蚜对吡虫啉敏感品系(LC₅₀为0.176 mg/L)和抗性品系(LC₅₀为14.657 mg/L)。采用阈性状分析方法,获得棉蚜对吡虫啉的抗性现实遗传力（h²）。【结果】相对于田间原始种群(LC₅₀为0.346 mg/L),吡虫啉敏感棉蚜品系对吡虫啉的LC50减少了2倍;获得的吡虫啉抗性棉蚜品系,经过40代的选育,得到抗性倍数为室内敏感品系的83.27倍的抗性品系。棉蚜对吡虫啉的抗性现实遗传力(h2)为0.1478。进一步预测其抗性发展速度,基于80%~90%的选择压力,预计抗性增长100倍时,吡虫啉可使用30.2~38.1代。【结论】这些研究说明棉蚜对吡虫啉存在抗性风险。     

Time-course analysis of the phenols in cucumber mosaic virus-resistant, -tolerant and -susceptible tomato genotypes

In this study, changes in quantity and quality of phenolic compounds were compared in cucumber mosaic virus (CMV)-inoculated and -un-inoculated plants of nine resistant, tolerant, susceptible and highly susceptible genotypes at three different time intervals. Total phenolic contents and the number of phenolic compounds were generally increased in CMV-inoculated plants. Maximum per cent increase in total phenolic contents over un-inoculated controls was observed as 77.55% in resistant genotype TMS-1, 84.17% in tolerant genotype L06238 and 82.88% in resistant genotype L02223 after 10, 20 and 30 days of inoculation, respectively. Thin layer chromatography of inoculated and un-inoculated plants indicates that in most of the tested genotypes, the number of phenolic compounds varied from cultivar to cultivar and within the same cultivar, depending upon the status of plants and growth stages. However, the trend of increase in quantity and quality of phenolic compounds in the tested units was not constant to draw a meaningful conclusion.