基于16S rDNA基因的中国部分地区非B型烟粉虱系统发育关系分析

【目的】烟粉虱Bemisia tabaci是一个快速进化的复合种。由于近年来烟粉虱危害不断增加, 其生物型问题也越来越受到关注。在我国, 不仅存在危害严重的B型烟粉虱, 同时也已发现多种非B型烟粉虱。通过鉴定我国部分地区目前发生的烟粉虱生物型种类, 分析它们与世界各地不同生物型之间的关系, 可为我国烟粉虱的生物型检测和综合防治提供依据。【方法】利用16S rDNA基因作为分子标记, 鉴定了2005年与2006年在我国6个省份（自治区）采集的22个非B型烟粉虱样本的生物型, 并探讨了各生物型之间的系统进化关系。【结果】所研究的22个非B型烟粉虱归属于Q型、Nauru型和An型, 3种生物型之间的遗传距离在10%以上, 但是Q生物型与B生物型之间的亲缘关系最近, 遗传距离在2.8%~4.0%范围内; 进化分歧数据还表明, 不同生物型之间的遗传距离明显大于同一生物型内遗传距离, 其中Q型内部差异最小, 在0.9%以内; 同时结果表明目前在我国多个生物型共同存在是一个普遍的现象, 其中Nauru型的分布较广泛; 在云南地区检测到的非B型烟粉虱生物型类型最多。【结论】烟粉虱生物型遗传分化复杂, 利用16S rDNA基因能有效鉴定烟粉虱的生物型。     

害虫生物型及其遗传机理

以农作物上5种重要害虫的生物型为例,简单地概括了分子标记在害虫生物型研究中的应用,重点阐述几种重要害虫如黑森瘿蚊,麦二叉蚜,褐飞虱,稻瘿蚊和烟粉虱等的生物型遗传变异机理。黑森瘿蚊、麦二叉蚜和稻瘿蚊的生物型由单基因控制,特定害虫生物型与作物基因型之间存在“基因_基因”关系;褐飞虱的致害性为多基因控制的数量性状;烟粉虱生物型的遗传变异机理尚不明确。还指出生物型遗传研究中待进一步解决的问题如生物型的遗传组成差异与致害力的关系等。     

环境因子对褐飞虱两种生物型种群参数影响的比较

比较了孟加拉型和生物型Ⅱ褐飞虱种群对高温、饥饿、浸水和杀虫剂等因子的反应。结果表明,35.0℃下孟加拉型若虫的致死中量时间(LT50)显著高于生物型Ⅱ,但两个生物型在26℃和37.5℃下的LT50间差异均不显著,而40.0℃时生物型Ⅱ若虫的LT50显著高于孟加拉型。在26、35.0℃和室内自然变温(26~35.0℃)条件下测定成虫耐饥力,两生物型间LT50均差异不显著。在各饥饿处理时间下,两生物型褐飞虱单雌平均产卵量差异均不显著。褐飞虱卵浸水1、3d和5d,两个生物型孵化率无显著差异,但生物型Ⅱ卵浸水5d后孵化率与对照相比差异显著,说明浸水5d对褐飞虱生物型Ⅱ卵的孵化率有影响。取食经浸水5d处理的稻株168h后,生物型Ⅱ若虫的死亡率与对照有显著差异,而孟加拉型死亡率与对照相比差异不显著;不浸水对照处理两个生物型的2~3龄若虫168h时就已全部发育为成虫,而浸水处理则到216h时才全部发育为成虫,这表明水稻浸水处理延迟了两个生物型若虫的发育进度。对于甲胺磷,药后1d、2d孟加拉型的致死中浓度(LC50)均显著小于生物型Ⅱ;对于噻嗪酮,孟加拉型药后5d的LC50均显著小于生物型Ⅱ,表明孟加拉型对这两种杀虫剂较生物型Ⅱ敏感。     

褐飞虱生物型研究进展:致害性变异的遗传机制

能够“克服”水稻抗性的褐飞虱生物型的出现 ,是目前水稻育种及害虫防治工作中面临的重大难题。为探明褐飞虱生物型的形成机制 ,许多学者致力于生物型致害性的遗传学研究 ,然而得出的结论却不尽一致。该文综述了有关褐飞虱生物型研究中争论较多的 3个方面 ,即生物型致害性的稳定性 ,致害性的遗传机制 ,以及生物型的形成机制 ,并介绍了最新的研究进展 ,展望了该领域的研究方向     

两种体色生物型桃蚜对杀虫剂敏感性差异及其与酶活力的关系

为了明确不同体色生物型桃蚜Myzus persicae (Suizer)对药剂的敏感性差异,本文采用室内生物测定和酶活力测定法检测不同体色生物型桃蚜对杀虫剂的敏感性以及与3种解毒酶和1种靶标酶活力关系。结果表明,红色生物型的敏感性明显低于绿色生物型,其中对吡虫啉的敏感性差异最大,LC50分别为6796648和284597 mg/L,红色为绿色生物型2388倍;其次为毒死蜱,LC50分别为12295798和1936816 mg/L,红色为绿色生物型635倍;两种生物型桃蚜对阿维菌素敏感性差异最小,红、绿色生物型的LC50分别为311678 和290966 mg/L,红色为绿色生物型107倍。红色生物型体内3种解毒酶（羧酸酯酶、谷胱甘肽-S-转移酶和多功能氧化酶P450）的比活力均高于绿色生物型,红色型羧酸酯酶比活力为绿色型的31倍,谷胱甘肽-S-转移酶为41倍,多功能氧化酶P450为15倍,两种体色生物型3种解毒酶的活力差异均达显著水平。两种体色生物型体内乙酰胆碱酯酶比活力没有差异,说明乙酰胆碱酯酶比活力与敏感性关系不大。     

mtCOI PCR-RFLP技术鉴别中国入侵型和土著型烟粉虱种群的有效性分析

烟粉虱Bemisia tabaci (Gennadius)是世界范围内最重要的入侵生物之一,准确地鉴别烟粉虱入侵生物型和土著生物型具有十分重要的现实意义。mtCOI PCR-RFLP技术具有快速、高效的特点,是当前应用最广泛的烟粉虱生物型鉴定技术,但是,不同限制性内切酶为基础的mtCOI PCR-RFLP技术在鉴定我国烟粉虱种群中的有效性仍不明了。本文比较了已报导的5种mtCOI PCR-RFLP技术在鉴别中国烟粉虱种群入侵生物型和土著生物型(B型、Q型,ZHJ1型、ZHJ2型、ZHJ3型)的有效性。结果表明,内切酶AluI不能区分B型和ZHJ2型烟粉虱;内切酶TaqI不能准确区分ZHJ3型和Q型烟粉虱个体;而内切酶VspI不仅不能准确区分ZHJ1型和Q型烟粉虱个体,也不能准确区分B型和ZHJ2型烟粉虱;内切酶MseI和Tru9I则不能有效鉴别上述5种烟粉虱生物型,因此不适宜推广使用。     

褐飞虱生物型研究中几个问题的商榷

目前在褐飞虱生物型监测中 ,由于鉴定品种及监测方法的不统一 ,使结果有出入 ,作者建议在对生物型 1进行监测时 ,鉴定品种应选用Mudgo[5] 。飞虱种群生物型的监测 ,宜以苗鉴定法为主 ,生存率测定法作参考。飞虱个体生物型测定以生存率法为主 ,苗鉴法为辅     

烟粉虱生物型研究进展

烟粉虱B em isia tabaci(G ennad ius)是一种取食植物汁液的重要农业害虫,同时也是许多植物病毒最重要的传播介体之一。烟粉虱被认为是由许多具有明显遗传分化的不同种群即生物型组成的复合种,其中B型烟粉虱是一种入侵性最强的生物型,几乎在世界各地均有分布。由于不同生物型的烟粉虱在寄主范围、传毒能力、地理分布、抗药性等许多生物学方面存在差异,因此对烟粉虱生物型的鉴定及其遗传分化研究对于该害虫的可持续控制具有重要的指导意义。烟粉虱生物型的鉴定方法包括生物学鉴定、酶谱鉴定以及分子标记鉴定的方法,其中使用的分子标记包括RAPD、AFLP、rDNA-ITS1、m tDNA CO I以及SSR等标记。目前,不同生物型的烟粉虱尤其B型烟粉虱的分类地位仍然存在争议。介绍了不同生物型烟粉虱的生物学差异、鉴定方法及其遗传分化研究的最新进展,同时探讨了不同生物型烟粉虱的分类地位和研究方向。     

油菜田看麦娘对高效氟吡甲禾灵产生抗药性的生理响应

采用种子检测法,鉴定了对除草剂高效氟吡甲禾灵产生抗药性的看麦娘(Alopecu-rus aequalis Sobol.)生物型JXRII。为探讨看麦娘对高效氟吡甲禾灵产生抗性的生理反应,以敏感生物型JJSII为对比,喷施高效氟吡甲禾灵后,测定抗药生物型和敏感生物型几个重要生理指标变化情况的差异。结果表明:高效氟吡甲禾灵处理下,JJSII的叶片电解质泄漏率、MDA含量极显著上升,叶绿素、类胡萝卜素含量下降;JXRII叶片的电解质渗漏率、MDA含量、叶绿素、类胡萝卜素均与对照无显著差异;高效氟吡甲禾灵处理下,JJSII的GSH含量在处理后就开始逐渐下降,JXRII的GSH含量始终高于对照;二者可溶性糖含量均增加,敏感生物型JJSII可溶性糖含量增加的幅度远远大于抗性生物型JXRII;这2种群体经除草剂胁迫下,除草剂对敏感性生物型的生理变化影响大,抗性生物型表现为一定的生理适应性。     

4种土壤调理剂对茶与枇杷间作茶园土壤改良效果研究

通过田间施肥实验研究了钙镁型、硅钙型、微生物型和生物炭型土壤调理剂对茶果间作茶园酸化土壤理化性质的影响。结果表明:钙镁型、硅钙型和生物炭型土壤调理剂,均可不同程度的提高土壤p H,微生物型土壤调理剂对土壤p H的影响不明显;生物炭型及含生物炭的复合配方可使土壤容重降低,土壤孔隙度增加,并能够显著提高土壤有机质的含量;通过土壤调剂的施用提高了枇杷的产量。     

不同种型布鲁菌间的基因获得与缺失研究

目的:了解不同种型布鲁菌间的基因差异及基因的获得与缺失情况。方法:采用生物信息学方法比较分析已测序的10株布鲁菌基因水平的差异,分析它们的核心基因组与泛基因组,对得到的差异基因用PCR验证其在19株不同生物型标准菌株中的分布情况。结果:不同种型布鲁菌在基因水平上存在较大差异,差异基因主要位于Ⅱ号染色体上;根据差异基因,鉴定了42个差异区段,这些差异区段在19株不同生物型标准菌株中存在差异分布。结论:布鲁菌在进化过程中分别获得或失去了不同的基因区段,从而适应不同的宿主环境。     

Characterization of intermediate biotypes in atrazine-susceptible populations of Chenopodium polyspermum L. and Amaranthus bouchonii thell. in Hungary

Intermediate biotypes for atrazine herbicide resistance in Chenopodium polyspermum and Amaranthus bouchonii were characterized by a peculiar chlorophyll fluorescence induction curve. The intermediate biotypes were isolated from progenies of susceptible plants in maize grown in alternate years without atrazine. The lethal dose in seedling treatments was lower than that of the resistant plants but higher than for susceptible plants. Atrazine at 10 μM was near the I₅₀-value for in vivo nitrite reductase activity in both intermediate biotypes. The activity of nitrite reducttase in the intermediate biotypes was about 75% of that of susceptible biotypes. These characteristics of intermediate biotypes were maternally inherited in crosses.     

Lipid composition of chloroplast membranes from weed biotypes differentially sensitive to triazine herbicides

Chloroplasts were isolated from triazine-sensitive and triazine-resistant biotypes of common groundsel (Senecio vulgaris L.), common lambsquarter (Chenopodium album L.), and redroot pigweed (Amaranthus retroflexus L.). Chloroplast lipids were extracted and analyzed for differences among sensitive and resistant biotypes. The distribution of lipid between major lipid classes differed in chloroplasts from resistant and susceptible biotypes. Chloroplasts from resistant biotypes contained higher proportions of monogalactosyl diglyceride and phosphatidyl ethanolamine and lower proportions of digalactosyl diglyceride and phosphatidyl choline than did chloroplasts from susceptible biotypes. Monogalactosyl diglyceride and phosphatidyl ethanolamine were also quantitatively higher in membranes of resistant versus susceptible biotypes. The major lipid classes of resistant chloroplast membranes contained lipids comparatively richer in unsaturated fatty acids with the exceptions of digalactosyl diglyceride from all three biotypes and phosphatidyl ethanolamine from common groundsel. Results correlated changes in triazine sensitivity with qualitative and quantitative differences in the lipid composition of chloroplast membranes.     

寄主型和迁飞型棉蚜的交配行为

棉蚜Aphis gossypii Glover种群存在寄主利用和迁飞能力上的明显分化,有棉花型和瓜型、迁飞型和滞留型之分。但是,棉花型与瓜型之间,以及迁飞型与滞留型之间是否能发生交配行为,尚无研究报道。本文在低温和短光照条件下分别诱导棉花型、瓜型、迁飞型和滞留型棉蚜的性蚜,并进行性蚜间的交配行为观察。结果表明,迁飞型和滞留型性蚜间可以发生交配行为,杂交时发生交配行为的个体比率与自交时无显著差异,但是杂交时雄蚜寻找配偶所花的时间要显著长于自交时,并且迁飞型雌蚜(M♀)与滞留型雄蚜(S♂)杂交时的交配持续时间也显著长于自交时。迁飞型和滞留型棉蚜同型交配容易完成,棉蚜的迁飞型和滞留型已在交配行为上产生了一定的分化。棉花型和瓜型棉蚜的性蚜间可以发生交配行为,并且两品系在正交和反交时雄蚜的寻偶时间与交配持续时间均无显著差异。     

微卫星位点BEM06与BEM23鉴别烟粉虱B型与Q型的有效性

烟粉虱Bemisia tabaci (Gennadius) B型与Q型是烟粉虱复合种中入侵性较强、分布较广的2种生物型, 当前在许多地区混合发生。这2种生物型的快速鉴别对其种群动态调查及入侵生态学研究具有重要价值。为了验证微卫星位点BEM06与BEM23鉴别B型与Q型烟粉虱的有效性, 本研究分析了这2个微卫星位点的等位基因在国内外17个B型、4个Q型、3个非B/Q型烟粉虱种群的分布特点。结果表明: 这2个微卫星位点的联合使用可鉴别B型与Q型烟粉虱, 但是无法有效地将B型、Q型与一些非B/Q型烟粉虱某些个体区分开来。结果提示: 利用微卫星位点BEM06与BEM23鉴别B型与Q型烟粉虱具有一定的局限性, 尤其是田间烟粉虱存在其他生物型时需要慎重使用这种鉴别方法。     

Relationship between saprotrophic growth in soil of different biotypes of Pochonia chlamydosporia and the infection of nematode eggs

The ecology of Pochonia chlamydosporia in soil and its interaction with both plant and nematode hosts are important for the successful exploitation of the fungus as a biological control agent. Differences in saprotrophism and parasitism were assessed for biotypes of P. chlamydosporia, which had originated from the eggs of cyst or root‐knot nematodes. Colonisation in soils of different textures (compost, sandy loam and loamy sand) measured by the numbers of colony‐forming units, differed greatly. Most biotypes were more abundant in sterilised soil of the different textures compared with non‐sterilised soils. The proportion of nematode eggs parasitised in a baiting technique demonstrated that biotypes had host preferences. Those biotypes that originated from root‐knot nematodes (RKN‐biotypes) infected significantly more Meloidogyne hapla eggs than Globodera pallida eggs, whereas biotypes from cyst nematodes (CN‐biotypes) parasitised more G. pallida eggs than M. hapla eggs. Differences in virulence between biotypes in an in vitro assay in which the fungi were placed directly onto the egg masses of M. hapla and those differences observed in the baiting technique showed similar trends. There was a negative linear correlation between the growth of the eight biotypes in soil and the proportion of eggs they infected in compatible interactions (i.e. fungal biotype originated from the same nematode genus as the target eggs). Those biotypes that infected most nematode eggs colonised soil the least extensively, suggesting that virulence may have a fitness cost. However, the relationship between saprotrophic growth and virulence is complex. The relative abundance of the different biotypes in soil in Petri dish assays was similar to that under glasshouse conditions using potato but not tomato as the plant host. Chlamydospores of some biotypes applied to soil significantly reduced (>50%) the population densities of M. hapla on tomato and of G. pallida on potato plants. Some biotypes that were both effective and virulent are good candidates for biological control of specific nematode pests. Data presented here and elsewhere indicate that RKN‐biotypes have different host preferences to CN‐biotypes; the specific primers based on the vcp1 gene from P. chlamydosporia rapidly confirmed the host origin of seven of the eight biotypes.     

Deoxyribonucleic acid reassociation in the classification of coagulase-positive staphylococci

DNA-DNA reassociation studies were performed with coagulase-positive staphylococci belonging to the biotypes A, B, C, D, E and F. These studies present genetic evidence for the existence of at least two distinct species within this group of organisms. The common Staphylococcus aureus strains were represented by organisms from biotypes A to D, and their DNA revealed over 80% nucleotide sequence homology under restrictive conditions. Less than 15% DNA homology was detected between strains from biotypes A to D (S. aureus) and those from biotypes E and F. The DNA of organisms from either the biotypes E or F displayed over 70% homology. Together, both biotypes are considered to represent the species S. intermedius. However, DNA homology values dropped to 50–65% between strains from different biotypes. This may justify the separation of S. intermedius biotypes E and F on a subspecies level.Abbreviations O.D. optical density - SSC standard saline citrate buffer (0.15 M NaCl, 0.015 M sodium citrate, pH 7.0) This work was supported by Deutsche Forschungsgemeinschaft     

Identification of Aphis gossypii Glover (Hemiptera: Aphididae) Biotypes from Different Host Plants in North China

Background

The cotton-melon aphid, Aphis gossypii Glover (Hemiptera: Aphididae), is a polyphagous species with a worldwide distribution and a variety of biotypes. North China is a traditional agricultural area with abundant winter and summer hosts of A. gossypii. While the life cycles of A. gossypii on different plants have been well studied, those of the biotypes of North China are still unclear.

Results

Host transfer experiments showed that A. gossypii from North China has two host-specialized biotypes: cotton and cucumber. Based on complete mitochondrial sequences, we identified a molecular marker with five single-nucleotide polymorphisms to distinguish the biotypes. Using this marker, a large-scale study of biotypes on primary winter and summer hosts was conducted. All A. gossypii collected from three primary hosts—hibiscus, pomegranate, and Chinese prickly ash—were cotton biotypes, with more cotton-melon aphids found on hibiscus than the other two species. In May, alate cotton and cucumber biotypes coexisted on cotton and cucumber seedlings, but each preferred its natal host. Both biotypes existed on zucchini, although the cucumber biotype was more numerous. Aphids on muskmelon were all cucumber biotypes, whereas most aphids on kidney bean were cotton biotypes. Aphids on seedlings of potato and cowpea belong to other species. In August, aphids on cotton and cucumber were the respective biotypes, with zucchini still hosting both biotypes as before. Thus, the biotypes had different fitnesses on different host plants.

Conclusions

Two host-specialized biotypes (cotton and cucumber) are present in North China. Hibiscus, pomegranate, and Chinese prickly ash can serve as winter hosts for the cotton biotype but not the cucumber biotype in North China. The fitnesses of the two host-specialized biotypes differ on various summer hosts. When alate aphids migrate to summer hosts, they cannot accurately land on the corresponding plant.     

Ultrastructural alterations to chloroplasts in triazine-resistant weed biotypes

Ultrastructural, morphometric and physiological techniques were used to determine the consistent chloroplast differences between triazine-resistant (R) and triazine-susceptible (S) biotypes of Amaranthus hybridus L., Chenopodium album L., and Brassica campestris L. All R biotypes had a larger proportion of the chloroplast volume as grana lamellae and a lower proportion of starch and stroma lamellae than S biotypes. In the R biotypes, a greater percentage of grana contain larger numbers of thylakoids per granum. A greater proportion of chlorophyll associated with the light-harvesting chlorophyll alb protein and a lower chlorophyll alb ratio, traits associated with an increase in grana lamellae, were noted in R biotypes. Chloroplasts of S biotypes could be modified to ultrastructural phenocopies of those in R biotypes by treatment with sublethal levels of the PSII inhibiting herbicides, bentazon, diuron, atrazine and prometon. Despite the structural similarities to R biotypes, the modified S biotypes were not resistant to atrazine as determined by fluorescence measurements. Thus, the structural alterations observed are apparently secondary effects of impaired photosynthetic electron transport in R biotypes, and are not the cause of triazine resistance.