Genetic diversity and geographic distribution of Bemisia tabaci and Trialeurodes vaporariorum in Costa Rica

The tobacco whitefly Bemisia tabaci (Gennadius) cryptic species complex and of the greenhouse whitefly Trialeurodes vaporariorum (Westwood) are extensively reported as destructive pests in vegetable crops worldwide. A survey was conducted in 2011 and 2012 to determine the occurrence and genetic diversity present in the populations of these whiteflies in the major vegetable production areas of Costa Rica. Insect samples were collected from sweet pepper (Capsicum annuum L.), tomato (Solanum lycopersicum L.), common bean (Phaseolus vulgaris L.) and weeds present in commercial crops either in open field or greenhouse conditions. PCR‐RFLP analysis of mitochondrial cytochrome c oxidase subunit 1 gene (mtCOI) sequences of 621 whitefly individuals confirmed the presence of the Mediterranean (MED) type of the B. tabaci and of T. vaporariorum in most sampled regions. Also, individuals of the Middle East‐Asia Minor 1 (MEAM1) type of the B. tabaci were observed in low numbers. Contingency analyses based on type of crop, geographical region, whitefly species, year of collection and production system confirmed that T. vaporariorum was the most frequent species in vegetable production areas in Costa Rica, both in greenhouses and in open fields. B. tabaci MED is likely spreading to new areas of the country, whereas B. tabaci MEAM1 was mostly absent or rarely found. Comparisons of mtCOI sequences from B. tabaci individuals revealed the presence of four B. tabaci sequence haplotypes (named MED‐i, MED‐ii, MEAM1‐i, MEAM1‐xviii) in Costa Rica, three of them identical to B. tabaci haplotypes previously reported in the Western Hemisphere and other parts of the world. Analysis of sequences of T. vaporariorum individuals revealed a more complex population with the presence of 11 haplotypes, two of which were identical to T. vaporariorum sequences reported from other countries.     

A multiple PCR‐LDR assay for identification of two invasive cryptic species of whiteflies Bemisia tabaci

The MEAM1 and MED species of the cryptic species complex Bemisia tabaci are important invasive pests that cause tremendous crop losses worldwide. A rapid and highly reliable molecular technique is necessary to identify these species because they are morphologically indistinguishable. Therefore, a multiple polymerase chain reaction coupled with a ligase detection reaction (PCR‐LDR) that was based on polymorphisms in the mitochondrial cytochrome oxidase I (mtCOI) gene of B. tabaci was developed to distinguish the two cryptic species. An assessment of the method indicated that PCR‐LDR provided high specificity and sensitivity in discriminating MEAM1 (SHB) and MED (SHQ) whiteflies. In field tests, PCR‐LDR genotyping was performed in one 96‐well plate to identify 93 individuals collected from 8 districts in the suburbs of Shanghai. Complete concordance was observed between PCR‐LDR and sequencing methods. The method was used to confirm that MEAM1 and MED were found in two districts, but only the MED was found in the other six districts. PCR‐LDR, which is a transplantable platform, provides an alternative method for species identification of B. tabaci at low cost.     

Species-specific COI primers for rapid identification of Bemisia tabaci Mediterranean (MED) species

Bemisia tabaci (Gennadius) is a rapidly evolving species complex, and is small in size and difficult to identify quickly and accurately. For the accurate identification and effective prevention of this species, the specific PCR method based on the mitochondrial DNA cytochrome oxidase subunit I (mt DNA COI) gene was used in the present study to evaluate rapid molecular detection technological applications for Mediterranean (MED) species. The MED was targeted and whitefly species from different regions were used as references. Fragments of the mt DNA COI gene of the MED and other closely related species were amplified with universal primers. Species-specific mitochondrial DNA cytochrome oxidase subunit I (SS-COI) primers BQLF/BQLR and BQJF/BQJR were designed from variable sites of MED and other whitefly species partial COI gene sequences. Subsequently, the lengths of target fragments were amplified by two pairs of SS-COI primers. Meanwhile, the accuracy, specificity and sensitivity of SS-COI primers were determined using various life stages of the MED and other related species collected from different locations. The primer pairs BQLF/BQLR and BQJF/BQJR generated 334 bp and 483 bp amplified fragment length respectively. Accuracy test results showed that primers can detect the MED single-head adults and also accurately detect single-egg and first instar, second instar and third instar nymphs, MED pupae, etc. Specific detection results demonstrated that the primers were able to amplify the MED but not the following species/populations: Middle East-Asia Minor 1 (MEAM1), Asia I, Asia II 1, Asia II 6 and Asia II 7, Aleurocanthus spiniferus (Quaintanca), A. camelliae, Siphoninus phillyreae, Aleuroclava rhododendri, A. thysanospermi, Aleurolobus taonabae, Dialeurodes citri and Trialeurodes vaporariorum (Westwood) in different areas. Sensitivity detection results showed that primers can detect the minimum threshold of 2,160 pg/μl and 1.38 pg/μl, respectively (equivalent to 1/1280 and 1/2000000 adult). This technique solves the problem that MED cannot be identified based on morphology. This method simultaneously adopted SS-COI PCR technological applications that improved detection accuracy and saved detection time.     

Chromosome‐level genome assembly of the greenhouse whitefly (Trialeurodes vaporariorum Westwood)

The greenhouse whitefly, Trialeurodes vaporariorum Westwood, is an agricultural pest of global importance. Here we report a 787‐Mb high‐quality draft genome sequence of T. vaporariorum assembled from PacBio long reads and Hi‐C chromatin interaction maps, which has scaffold and contig N50 lengths of 70 Mb and 500 kb, respectively, and contains 18,275 protein‐coding genes. About 98.8% of the assembled contigs were placed onto the 11 T. vaporariorum chromosomes. Comparative genomic analysis reveals significantly expanded gene families such as aspartyl proteases in T. vaporariorum compared to Bemisia tabaci Mediterranean (MED) and Middle East‐Asia Minor 1 (MEAM1). Furthermore, the cytochrome CYP6 subfamily shows significant expansion in T. vaporariorum and several genes in this subfamily display developmental stage‐specific expression patterns. The high‐quality T. vaporariorum genome provides a valuable resource for research in a broad range of areas such as fundamental molecular ecology, insect–plant/insect–microorganism or virus interactions and pest resistance management.     

Genetic groups and endosymbiotic microbiota of the Bemisia tabaci species complex in Japanese agricultural sites

The whitefly Bemisia tabaci is a cryptic species complex of at least 24 genetically distinct species. Thus far, one obligate and seven facultative symbiotic bacteria have been reported from the B. tabaci species complex. Both genetic groups and infected symbionts are extremely important to estimate the pest status of B. tabaci. In this study, we collected 340 whiteflies from 39 agricultural sites, covering an entire region of the B. tabaci habitat in Japan, and examined the genotypes and symbiont community composition at subspecies level. Use of the cleaved amplified polymorphic sequence technique and mitochondrial cytochrome oxidase subunit I gene sequencing detected five genetic groups: indigenous species JpL and Asia II 6, invasive species Middle East‐Asia Minor 1 (MEAM1) and Mediterranean Subclade Q1 (MED Q1), and a genetic group previously undetected in Asia, Mediterranean Subclade Q2 (MED Q2). The genetic groups exhibited characteristic infection statuses with regard to their facultative symbionts, as observed in other countries. The endosymbiotic microbiota of the Japanese MED Q1 was different from that in neighbouring countries, but similar to that in the French or Uruguayan MED Q1. These results may indicate that Japanese MED Q1 species have not invaded from neighbouring countries, but from distant countries by international transportation. All Japanese MED Q2 species were infected with Rickettsia, some of which are regarded as conferring a female‐biased sex ratio and fitness benefit on B. tabaci. The results suggest that MED Q2 may be prevalent in Japan and neighbouring countries.     

Genome-wide identification and analysis of sulfatase and sulfatase modifying factor genes in Bemisia tabaci (Hemiptera: Aleyrodidae)

The invasive pest whitefly (Bemisia tabaci) is a complex species, of which Middle East-Minor Asia 1 (MEAM1) and Mediterranean (MED) are the two most damaging members. Previous research showed that cabbage is frequently infested with MEAM1 but seldomly with MED, and this difference in performance is associated with glucosinolate (GS) content. Some insects can modify GS using glucosinolate sulfatase (SULF), the activity of which is regulated by sulfatase modifying factor 1 (SUMF1); therefore, to increase our understanding of different performances of MEAM1 and MED on cabbage plants, we identified and compared nine putative SULFs and one SUMF in MEAM1 and MED. We found that the lengths of two genes, BtSulf2 and BtSulf4, differed between MEAM1 and MED. The messenger RNA levels of BtSulf4 increased more than 20-fold after MEAM1 and MED adults were exposed to GS, but BtSulf2 expression was only induced by GS in MEAM1. Knockdown of BtSulf2 and BtSulf4 in MEAM1 resulted in a substantial increase in the mortality of GS-treated adults but not in MED. These results indicate that differences in BtSulf2 and BtSulf4 sequences and/or expression may explain why MEAM1 performs better than MED on cabbage. Our results provide a basis for future functional research on SULF and SUMF in B. tabaci.     

A new record for Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) species complex of Turkey

In this study, species complex of Turkish Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) populations was determined by PCR‐based DNA analysis. According to phylogenetic analyses, the B. tabaci samples have been identified within three generic groups. A major part of the samples belonged to two invasive species, either Middle East–Asia Minor 1 (MEAM1) or Mediterranean (MED). In addition to these two invasive species, several samples collected from greenhouses and cotton fields have been found to be related to Middle East–Asia Minor 2 (MEAM2), which is the first record of Turkish B. tabaci species complex.     

The ability to manipulate plant glucosinolates and nutrients explains the better performance of Bemisia tabaci Middle East‐Asia Minor 1 than Mediterranean on cabbage plants

The performance of herbivorous insects is greatly affected by host chemical defenses and nutritional quality. Some herbivores have developed the ability to manipulate plant defenses via signaling pathways. It is currently unclear, however, whether a herbivore can benefit by simultaneously reducing plant defenses and enhancing plant nutritional quality. Here, we show that the better performance of the whitefly Bemisia tabaci Middle East‐Asia Minor 1 (MEAM1; formerly the “B” biotype) than Mediterranean (MED; formerly the “Q” biotype) on cabbage is associated with a suppression of glucosinolate (GS) content and an increase in amino acid supply in MEAM1‐infested cabbage compared with MED‐infested cabbage. MEAM1 had higher survival, higher fecundity, higher intrinsic rate of increase (r_m), a longer life span, and a shorter developmental time than MED on cabbage plants. Amino acid content was higher in cabbage infested with MEAM1 than MED. Although infestation by either biotype decreased the levels of total GS, aliphatic GS, glucoiberin, sinigrin, glucobrassicin, and 4OH‐glucobrassicin, and the expression of related genes in cabbage, MED infestation increased the levels of 4ME‐glucobrassicin, neoglucobrassicin, progoitrin, and glucoraphanin. The GS content and expression of GS‐related genes were higher in cabbage infested with MED than with MEAM1. Our results suggest that MEAM1 performs better than MED on cabbage by manipulating host defenses and nutritional quality.     

Phenolics,rather than glucosinolates,mediate host choice of Bemisia tabaci MEAM1 and MED on five cabbage genotypes

Worldwide, the most two important cryptic species of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) are MEAM1 (Middle East–Asia Minor 1, “B” biotype) and MED (Mediterranean, “Q” biotype). Although both B. tabaci MEAM1 and MED are polyphagous, they differ greatly in host choice and performance on various host plants. MEAM1 prefer to settle and perform better than MED on cabbage (Brassica oleracea), for example, but the underlying mechanism is largely unexplored. In the present study, we first measured the contents of the main secondary insect-resistant substances (glucosinolates and phenolics) and main nutrients (soluble proteins, total amino acids and total nitrogen) in five cabbage genotypes. We then examined the settling and oviposition choices of MEAM1 and MED on the five cabbage genotypes, respectively. The settling and oviposition preferences of both MEAM1 and MED were negatively related to the content of total phenolics rather than to the content of glucosinolates or main nutrients. Furthermore, our results showed that MEAM1 ranked the host quality of the cabbage genotypes more accurately than MED. The results at least in part indicate that total phenolics rather than glucosinolates mediate the host choice of B. tabaci MEAM1 and MED on the five cabbage genotypes.     

利用TaqMan等位基因技术鉴定烟粉虱MEAM1和MED隐种

MEAM1和MED是烟粉虱Bemisia tabaci两种重要的外来入侵隐种, 在我国部分地区常混合发生, 对我国农业生产造成了不同程度的危害和损失。尤其是MED隐种危害寄主范围更广, 对多种杀虫剂具有较高抗性, 防治上更为困难。因此, 如何快速鉴定烟粉虱MEAM1和MED隐种, 对于烟粉虱防治策略的选择具有十分重要的意义。本研究选择线粒体细胞色素氧化酶I(mitochondrial cytochrome oxidase I, mtDNA COI)基因保守区域内的单核苷酸多态性（SNP）为靶标, 应用等位基因聚合酶链式反应技术, 借助TaqMan-MGB荧光染色标记探针, 建立了一种鉴定烟粉虱MEAM1和MED隐种的等位基因选择性PCR方法, 并对北京11个区县的14个烟粉虱种群进行了隐种鉴定。结果表明, 北京地区14个烟粉虱种群样本与已知烟粉虱MED隐种种群在荧光值分布上聚为一簇, 为MED隐种。该鉴定方法具备SNP基因分型的优点, 可快速、 可靠、 高通量地鉴定烟粉虱MEAM1和MED, 为烟粉虱隐种鉴定及遗传分化研究提供了新的可选途径。     

Cloning of a putative extracellular Cu/Zn superoxide dismutase and functional differences of superoxide dismutases in invasive and indigenous whiteflies

Superoxide dismutases (SODs) are a group of important antioxidant defense enzymes. In this study, a putative extracellular Cu/Zn superoxide dismutase (ecCuZnSOD) complementary DNA was cloned and characterized from the whitefly, Bemisia tabaci. Quantitative polymerase chain reaction analysis showed that the expression level of Bt‐ecCuZnSOD was more than 10‐fold higher in the invasive Middle East Asia Minor 1 (MEAM1) than in the native Asia II 3 species of the B. tabaci species complex. After exposure to low temperature (4 °C), the expression of Bt‐ecCuZnSOD gene was significantly up‐regulated in MEAM1 but not in Asia II 3. Furthermore, the expression level of B. tabaci intracellular CuZnSOD (Bt‐icCuZnSOD), Bt‐ecCuZnSOD and mitochondrial MnSOD (Bt‐mMnSOD) was compared after transferring MEAM1 and Asia II 3 whiteflies from favorable (cotton) to unfavorable host plants (tobacco). On cotton, both CuZnSOD genes were expressed at a higher level in MEAM1 compared with Asia II 3. Interestingly, after transferring onto tobacco, the expression of Bt‐ecCuZnSOD was significantly induced in Asia II 3 but not in MEAM1. On the other hand, while Bt‐mMnSOD was expressed equally in both species on cotton, Bt‐mMnSOD messenger RNA was up‐regulated in MEAM1 on tobacco. Consistently, enzymatic activity assays of CuZnSOD and MnSOD demonstrated that CuZnSOD might play an important protective role against oxidative stress in Asia II 3, whereas MnSOD activation was critical for MEAM1 whiteflies during host adaptation. Taken together, our results suggest that the successful invasion of MEAM1 is correlated with its constitutive high activity of CuZnSOD and inducible expression of MnSOD under stress conditions.     

Abundance of Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) and its parasitoids on vegetables and cassava plants in Burkina Faso (West Africa)

The whitefly Bemisia tabaci is a pest of many agricultural and ornamental crops worldwide and particularly in Africa. It is a complex of cryptic species, which is extremely polyphagous with hundreds of host plants identified around the world. Previous surveys in western Africa indicated the presence of two biotypes of the invasive MED species (MED‐Q1 and MED‐Q3) living in sympatry with the African species SSA and ASL. This situation constitutes one of the rare cases of local coexistence of various genetic entities within the B. tabaci complex. In order to study the dynamics of the distribution and abundance of genetic entities within this community and to identify potential factors that could contribute to coexistence, we sampled B. tabaci populations in Burkina Faso in 2015 and 2016 on various plants, and also their parasitoids. All four genetic entities were still recorded, indicating no exclusion of local species by the MED species. While B. tabaci individuals were found on 55 plant species belonging to eighteen (18) families showing the high polyphagy of this pest, some species/biotypes exhibited higher specificity. Two parasitoid species (Eretmocerus mundus and Encarsia vandrieschei) were also recorded with E. mundus being predominant in most localities and on most plants. Our data indicated that whitefly abundance, diversity, and rate of parasitism varied according to areas, plants, and years, but that parasitism rate was globally highly correlated with whitefly abundance suggesting density dependence. Our results also suggest dynamic variation in the local diversity of B. tabaci species/biotypes from 1 year to the other, specifically with MED‐Q1 and ASL species. This work provides relevant information on the nature of plant–B. tabaci‐parasitoid interactions in West Africa and identifies that coexistence might be stabilized by niche differentiation for some genetic entities. However, MED‐Q1 and ASL show extensive niche overlap, which could ultimately lead to competitive exclusion.     

Infection density pattern of Cardinium affects the responses of bacterial communities in an invasive whitefly under heat conditions

Communities of bacteria, especially symbionts, are vital for the growth and development of insects and other arthropods, including Bemisia tabaci Mediterranean (MED), a destructive and invasive insect pest. However, the infection density patterns and influence factors of bacteria in whiteflies, which mainly include symbionts, remain largely unclear. To reveal the different density patterns of Cardinium in B. tabaci MED populations and the impacts of high temperatures on whiteflies with different Cardinium density infection patterns, 2 isofemale lines isolated from B. tabaci MED from the same geographical population of China and from B. tabaci MED collected from other countries and locations were examined using several techniques and methods, including fluorescence in situ hybridization (FISH), quantitative real-time polymerase chain reaction (qPCR), 16S rRNA gene sequencing, and 2b-RAD sequencing. The results showed that there were 2 different infection density patterns of Cardinium in B. tabaci MED (including 1 high-density pattern and 1 low-density pattern). For whiteflies with low-density Cardinium, conventional PCR could not detect Cardinium, but the other techniques confirmed that there was a low level of Cardinium within hosts. High temperature significantly decreased the diversity of bacterial communities: the relative titer of Cardinium increased but the density of Rickettsia decreased in the isofemale line with high-density Cardinium. However, high temperature did not influence the diversity and symbiont density in the line with low-density Cardinium. Moreover, high temperature influenced the functions of bacterial communities in whiteflies with high-density Cardinium but did not affect the bacterial functions in whiteflies with low-density Cardinium. Our results provide novel insights into the complex associations between symbionts and host insects.     

Patterns of host cell inheritance in the bacterial symbiosis of whiteflies

Whiteflies possess bacterial symbionts Candidatus Portiera aleyrodidium that are housed in specialized cells called bacteriocytes and are faithfully transmitted via the ovary to insect offspring. In one whitefly species studied previously, Bemisia tabaci MEAM1, transmission is mediated by somatic inheritance of bacteriocytes, with a single bacteriocyte transferred to each oocyte and persisting through embryogenesis to the next generation. Here, we investigate the mode of bacteriocyte transmission in two whitefly species, B. tabaci MED, the sister species of MEAM1, and the phylogenetically distant species Trialeurodes vaporariorum. Microsatellite analysis supported by microscopical studies demonstrates that B. tabaci MED bacteriocytes are genetically different from other somatic cells and persist through embryogenesis, as for MEAM1, but T. vaporariorum bacteriocytes are genetically identical to other somatic cells of the insect, likely mediated by the degradation of maternal bacteriocytes in the embryo. These two alternative modes of transmission provide a first demonstration among insect symbioses that the cellular processes underlying vertical transmission of bacterial symbionts can diversify among related host species associated with a single lineage of symbiotic bacteria.     

Identification,comparison, and functional analysis of salivary phenol‐oxidizing enzymes in Bemisia tabaci B and Trialeurodes vaporariorum

The differences in the ability of the invading whitefly, Bemisia tabaci (Gennadius) (commonly known as biotype B and hereafter as B) and Trialeurodes vaporariorum (Westwood) (both Hemiptera: Aleyrodidae) to utilize salivary phenol‐oxidizing enzymes – polyphenol oxidase (PPO) and peroxidase (POD) to detoxify plant defensive phenolic compounds were explored. Polyphenol oxidase and POD were found in the saliva of both B and T. vaporariorum. For tomato colonies, the PPO and POD activities in the watery saliva of B were 2.27‐ and 1.34‐fold higher than those of T. vaporariorum. The PPO activities against specific phenolic compounds commonly found in plants were compared. The activities of those from B were significantly greater than those from T. vaporariorum. We also measured PPO activity in both species after they had fed on plants that were undamaged or had been previously damaged with either a plant pathogen [Phytophthora infestans (Mont.) de Bary (Peronosporales)] infection, mechanical damage, B infestation, or exogenous salicylic acid. For B, PPO activities in watery saliva increased 229, 184, 152, and 139% in response to the four treatments, whereas those of T. vaporariorum only increased 133, 119, 113, and 103%, respectively. Biotype B infestation significantly increased the total phenolic content of tomato leaves. Meanwhile, feeding on tomato infestation with B had no significant effect on the survival rate of B, but decreased the survival rate of T. vaporariorum significantly. These results suggest that B has stronger ability utilizing PPO to detoxify high concentrations of phenolics than T. vaporariorum, and this contributes to a significant advantage for B to hold high fitness on plants with induced resistance. Possible roles of salivary PPO in the competition between B and T. vaporariorum are discussed.     

Effects of temperature on the growth and reproduction characteristics of Bemisia tabaci B‐biotype and Trialeurodes vaporariorum

The development period, survival rate, longevity and fecundity of two whiteflies, Bemisia tabaci B‐biotype and Trialeurodes vaporariorum (Homoptera: Aleyrodidae) were compared under different temperature laboratory conditions (15°C, 18°C, 21°C and 24°C). Egg development of B. tabaci B‐biotype was significantly longer compared with that of T. vaporariorum at 15°C, 18°C and 24°C. Significantly longer pseudo‐pupae development and lower survival rate were found in B. tabaci B‐biotype at 15°C compared with those at 18°C, 21°C and 24°C. Significantly higher fecundity was found in B. tabaci B‐biotype at 24°C compared with that at 15°C, 18°C and 21°C. However, the fecundity of T. vaporariorum was significantly lower at 24°C relative to that at 15°C, 18°C and 21°C. Significantly shorter 1st instar larval development was found in T. vaporariorum compared with that of B. tabaci at 15°C and 18°C. Significantly longer 2nd instar larval development was found in B. tabaci and T. vaporariorum at 15°C compared with that at 18°C, 21°C and 24°C. However, significantly shorter 3rd instar larval development was found in T. vaporariorum compared with that of B. tabaci at 15°C, 18°C and 24°C. The adaptive divergence of tolerance to relatively low temperature may be an important factor that results in the interspecific differentiation between the seasonal dynamics of these two whiteflies in China.     

A shift of vector specificity acquired by a begomovirus through natural homologous recombination

Recombination is common in plant viruses such as geminiviruses, but the ecological and pathogenic consequences have been explored only in a few cases. Here, we found that a new begomovirus, tomato yellow leaf curl Shuangbai virus (TYLCSbV), probably originated from the recombination of Ageratum yellow vein China virus (AYVCNV) and tobacco curl shoot virus (TbCSV). Agrobacterium-mediated inoculation showed that TYLCSbV and AYVCNV have similar levels of infectivity on tomato and tobacco plants. However, the two viruses exhibit contrasting specificities for vector transmission, that is, TYLCSbV was efficiently transmitted by the whitefly Bemisia tabaci Mediterranean (MED) rather than by the whitefly B. tabaci Middle East-Asia Minor 1 (MEAM1), whereas AYVCNV was more efficiently transmitted by MEAM1. We also showed that the transmission efficiencies of TYLCSbV and AYVCNV are positively correlated with the accumulation of the viruses in whitefly whole bodies and organs/tissues. The key coat protein amino acids that determine their accumulation are between positions 147 and 256. Moreover, field surveys suggest that MED has displaced MEAM1 in some regions where TYLCSbV was collected. Viral competition assays indicated that TYLCSbV outcompeted AYVCNV when transmitted by MED, while the outcome was the opposite when transmitted by MEAM1. Our findings suggest that recombination has resulted in a shift of vector specificity that could provide TYLCSbV with a potential selective transmission advantage, and the population shift of whitefly cryptic species could have influenced virus evolution towards an extended trajectory of transmission.     

烟粉虱的分类地位及在中国的分布

烟粉虱广泛分布于全球热带和亚热带地区。近20多年,烟粉虱的一些遗传群入侵世界各地,严重危害作物生产。烟粉虱遗传结构的多样性和复杂性早已被关注,但其分类地位,尤其是烟粉虱到底是一个包含多个生物型的种还是一个包含许多隐种的物种复合体,一直颇受争议。近几年,有关烟粉虱种系发生和系统学的研究取得长足进展,有证据推论其是一个包含至少31个隐种的物种复合体,但生殖隔离证据仍显不足,种系发生分析结果也因仅依据COI一个基因而受到质疑。因此,在大多数从事烟粉虱研究的同行接受其为一个物种复合体的概念的同时,仍有同行沿用生物型的概念。在我国境内已先后报道了包括13个本地种和2个全球入侵种在内的15个烟粉虱隐种。本地种主要分布在我国南部及包括海南岛和台湾岛的东南沿海地区,隐种的多样性由南向北逐渐降低。入侵种“中东一小亚细亚1”隐种（MEAMl）（即“B型”）和“地中海”隐种（MED）（即“Q型”）分别于20世纪90年代中后期和2003年前后入侵我国,并在许多地区迅速取代了本地种而占据优势地位。全国范围内的调查数据显示,这2个入侵种可在大部分区域共同存在,但自2005年以来,MED在许多地区陆续取代MEAMl,这很可能与MED对大量使用的新烟碱类杀虫剂有较强抗性有关。本文还讨论了烟粉虱隐种复合体分类所面临的命名等难题以及大范围抽样调查的数据偏差问题。     

Development of 28 EST‐SSR markers based on transcriptome sequences of Gobiobotia filifer and cross‐species amplification

Gobiobotia filifer is a small benthic fish distributed in Yangtze River Basin. The abundance of G. filifer increased after impoundment of Xiluodu Dam and Xiangjiaba Dam. The state of population structure and changes of genetic diversity before and after impoundment of Xiluodu Dam and Xiangjiaba Dam were interesting issues. However, efficient molecular markers were rare, which will limit us to solve above problems. Twenty‐eight expressed sequence tag SSRs (EST‐SSRs) were successfully identified and verified as stable amplification and polymorphic loci by polyacrylamide gel electrophoresis (PAGE) and capillary electrophoresis. The number of alleles at these EST‐SSR loci ranged from 3 to 14, the polymorphism information content values were 0.125–0.897, and the observed and expected heterozygosities were 0.0–0.857 and 0.132–0.928, respectively. Cross‐species amplification of the 28 loci developed in this study was examined in seven individuals of each of the 7 taxa. The amplification efficiency of 28 EST‐SSRs primer pairs is related to the distance of genetic relationship between cross‐species with G. filifer, and same subfamily species (Xenophysogobio boulengeri and Xenophysogobio nudicorpa) showed the highest (50%) amplification efficiency. These EST‐SSR markers could be used to analyse genetic diversity and population structure of G. filifer and related species.