Oral Delivery of Double-Stranded RNAs and siRNAs Induces RNAi Effects in the Potato/Tomato Psyllid, Bactericerca cockerelli
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{"title"=>"Oral delivery of double-stranded RNAs and siRNAs induces RNAi effects in the potato/tomato psyllid, Bactericerca cockerelli", "type"=>"journal", "authors"=>[{"first_name"=>"Hada", "last_name"=>"Wuriyanghan", "scopus_author_id"=>"22235941300"}, {"first_name"=>"Cristina", "last_name"=>"Rosa", "scopus_author_id"=>"22935820100"}, {"first_name"=>"Bryce W.", "last_name"=>"Falk", "scopus_author_id"=>"7101998497"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"362925708", "sgr"=>"81155134267", "issn"=>"19326203", "pmid"=>"22110747", "scopus"=>"2-s2.0-81155134267", "doi"=>"10.1371/journal.pone.0027736", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)"}, "id"=>"e77de20b-dff0-34fa-820f-025201a86a0f", "abstract"=>"The potato/tomato psyllid, Bactericerca cockerelli (B. cockerelli), and the Asian citrus psyllid, Diaphorina citri (D. citri), are very important plant pests, but they are also vectors of phloem-limited bacteria that are associated with two devastating plant diseases. B. cockerelli is the vector of Candidatus Liberibacter psyllaurous (solanacearum), which is associated with zebra chip disease of potatoes, and D. citri is the vector of Ca. Liberibacter asiaticus, which is associated with the Huanglongbing (citrus greening) disease that currently threatens the entire Florida citrus industry. Here we used EST sequence information from D. citri to identify potential targets for RNA interference in B. cockerelli. We targeted ubiquitously expressed and gut-abundant mRNAs via injection and oral acquisition of double-stranded RNAs and siRNAs and were able to induce mortality in recipient psyllids. We also showed knockdown of target mRNAs, and that oral acquisition resulted primarily in mRNA knockdown in the psyllid gut. Concurrent with gene knockdown was the accumulation of target specific ∼ 21 nucleotide siRNAs for an abundant mRNA for BC-Actin. These results showed that RNAi can be a powerful tool for gene function studies in psyllids, and give support for continued efforts for investigating RNAi approaches as possible tools for psyllid and plant disease control.", "link"=>"http://www.mendeley.com/research/oral-delivery-doublestranded-rnas-sirnas-induces-rnai-effects-potatotomato-psyllid-bactericerca-cock", "reader_count"=>89, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Librarian"=>2, "Researcher"=>23, "Student > Doctoral Student"=>10, "Student > Ph. D. Student"=>27, "Student > Postgraduate"=>2, "Student > Master"=>9, "Student > Bachelor"=>8, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>2, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Librarian"=>2, "Researcher"=>23, "Student > Doctoral Student"=>10, "Student > Ph. D. Student"=>27, "Student > Postgraduate"=>2, "Student > Master"=>9, "Student > Bachelor"=>8, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>2, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>5, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>78, "Medicine and Dentistry"=>1, "Chemistry"=>1, "Economics, Econometrics and Finance"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Economics, Econometrics and Finance"=>{"Economics, Econometrics and Finance"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>78}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>5}}, "reader_count_by_country"=>{"China"=>1, "Israel"=>1, "Slovenia"=>1}, "group_count"=>5}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/360818", "https://ndownloader.figshare.com/files/360827", "https://ndownloader.figshare.com/files/360835", "https://ndownloader.figshare.com/files/360841", "https://ndownloader.figshare.com/files/360852", "https://ndownloader.figshare.com/files/360867", "https://ndownloader.figshare.com/files/360872"], "description"=>"<div><p>The potato/tomato psyllid, <em>Bactericerca cockerelli</em> (<em>B. cockerelli</em>), and the Asian citrus psyllid, <em>Diaphorina citri</em> (<em>D. citri</em>), are very important plant pests, but they are also vectors of phloem-limited bacteria that are associated with two devastating plant diseases. <em>B. cockerelli</em> is the vector of <em>Candidatus</em> Liberibacter psyllaurous (solanacearum), which is associated with zebra chip disease of potatoes, and <em>D. citri</em> is the vector of <em>Ca</em>. Liberibacter asiaticus, which is associated with the Huanglongbing (citrus greening) disease that currently threatens the entire Florida citrus industry. Here we used EST sequence information from <em>D. citri</em> to identify potential targets for RNA interference in <em>B. cockerelli</em>. We targeted ubiquitously expressed and gut-abundant mRNAs via injection and oral acquisition of double-stranded RNAs and siRNAs and were able to induce mortality in recipient psyllids. We also showed knockdown of target mRNAs, and that oral acquisition resulted primarily in mRNA knockdown in the psyllid gut. Concurrent with gene knockdown was the accumulation of target specific ∼ 21 nucleotide siRNAs for an abundant mRNA for <em>BC-Actin</em>. These results showed that RNAi can be a powerful tool for gene function studies in psyllids, and give support for continued efforts for investigating RNAi approaches as possible tools for psyllid and plant disease control.</p> </div>", "links"=>[], "tags"=>["double-stranded", "rnas", "sirnas", "induces", "rnai", "effects"], "article_id"=>131329, "categories"=>["Molecular Biology", "Genetics", "Cell Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.s001", "https://dx.doi.org/10.1371/journal.pone.0027736.s002", "https://dx.doi.org/10.1371/journal.pone.0027736.s003", "https://dx.doi.org/10.1371/journal.pone.0027736.s004", "https://dx.doi.org/10.1371/journal.pone.0027736.s005", "https://dx.doi.org/10.1371/journal.pone.0027736.s006", "https://dx.doi.org/10.1371/journal.pone.0027736.s007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Oral_Delivery_of_Double_Stranded_RNAs_and_siRNAs_Induces_RNAi_Effects_in_the_Potato_Tomato_Psyllid_Bactericerca_cockerelli_/131329", "title"=>"Oral Delivery of Double-Stranded RNAs and siRNAs Induces RNAi Effects in the Potato/Tomato Psyllid, <em>Bactericerca cockerelli</em>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-11-16 00:22:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/712326"], "description"=>"<p>Total RNAs (1.5 µg) from <i>B. cockerelli (BC)</i> and <i>D. citri (DC)</i> were separated by denaturing agarose gel electrophoresis, transferred to nylon membranes and hybridized with <sup>32</sup>P-UTP-labeled probes prepared using corresponding <i>B. cockerelli</i> sequences. 0.5–10 Kb RNA ladder (Invitrogen) was used on the same gel and the sizes are indicated. Labels A – D above each panel indicate the specific probe used for the corresponding blot. The exposure times for <i>BC-Actin</i>, <i>BC-ATPase</i>, <i>BC-Hsp70</i> and <i>BC-CLIC</i> were 15 h, 2 h, 38 h and 38 h, respectively.</p>", "links"=>[], "tags"=>["blot", "analyses", "hsp70", "mrnas"], "article_id"=>382686, "categories"=>["Molecular Biology", "Genetics", "Plant Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Northern_blot_analyses_of_Actin_ATPase_Hsp70_and_CLIC_mRNAs_in_B_cockerelli_and_D_citri_/382686", "title"=>"Northern blot analyses of <i>Actin, ATPase, Hsp70 and CLIC</i> mRNAs in <i>B. cockerelli</i> and <i>D. citri</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-16 00:44:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/712423"], "description"=>"<p>Total RNAs were extracted from dissected <i>B. cockereli</i> tissues including gut, head, abdomen, thorax, and reproductive organ (RO). cDNA was synthesized and semi-quantitative RT-PCR analyses were performed on <i>BC-Actin</i>, <i>BC-ATPase</i>, <i>BC-Hsp70</i> and <i>BC-CLIC</i>. Expression profile of <i>rRNA</i> was used as control. RT-PCR was carried out 30 cycles for <i>BC-ATPase</i>, <i>BC-Hsp70</i> and <i>BC-CLIC</i>, and 22 cycles for <i>BC-Actin</i> and <i>rRNA</i> using GoTaq polymerase. The PCR reactions are: initial denaturation at 94°C for 5 min, followed by cycles with 94°C (30 s), 55°C (30 s), 72°C (1 min), and a final extension for 7 min at 72°C.</p>", "links"=>[], "tags"=>["accumulation", "mrnas"], "article_id"=>382787, "categories"=>["Molecular Biology", "Genetics", "Plant Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_relative_accumulation_of_B_cockerelli_mRNAs_in_different_tissues_/382787", "title"=>"The relative accumulation of <i>B. cockerelli</i> mRNAs in different tissues.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-16 00:46:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/712475"], "description"=>"<p>(A) Psyllid feeding system. Teneral pale-colored adult psyllids are placed in plastic vials (25mm X 45mm). The feeding solution is 15% sucrose (w: v) sandwiched between two sheets of stretched parafilm. Plastic vials containing the psyllids were maintained at room temperature in the dark. (B) Psyllids were fed on 15% sucrose containing 50ng/µL of GFP PCR product. After 48h feeding, psyllids were collected, DNA extracted and analyzed by PCR. Lane 1, 1Kb plus DNA Marker; Lane 2 and 3, 0.004ng and 0.0004 ng GFP PCR product only as a positive control for PCR reaction. Lanes 4 and 5, PCR products from GFP DNA-fed psyllids; Lane 6, PCR product from sucrose-fed psyllid extract mixed with 0.004 ng GFP DNA; Lane 7, sucrose-fed psyllid used as negative control. (C) 1∶10 (v:v) dilution of green coloring dye was added to 15% sucrose and used for feeding. After 24h the psyllids were examined by low power light microscopy. Upper row is food coloring-fed psyllids and the lower row shows sucrose-fed psyllid controls. (D) 50 ng/µL Cy™3-labeled GFP dsRNA was supplied in 15% sucrose and fed for psyllids. After 48h feeding, the fluorescence was visualized by fluorescence microscopy (Leica DM5000B) equipped with a mercury lamp (ebq100, ISOLATED). In each panel, the lower psyllid was fed with Cy™3-labeled dsRNA and the upper psyllid was sucrose-fed psyllid control. Psyllids were visualized under white light (a) or under fluorescent light of different excitations at 460nm, 480nm and 436nm (b-d).</p>", "links"=>[], "tags"=>["ingestion"], "article_id"=>382850, "categories"=>["Molecular Biology", "Genetics", "Plant Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Validation_of_oral_ingestion_for_B_cockerelli_/382850", "title"=>"Validation of oral ingestion for <i>B. cockerelli</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-16 00:47:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/712573"], "description"=>"<p>Groups of 30 teneral adult psyllids were used for each treatment. The experiments were repeated three times. 15% sucrose was used as a food control and H<sub>2</sub>O was used as a non-food control for psyllid mortality. GFP dsRNA was used as a dsRNA control. The Bonferroni (Dunn) t-test was used for statistical analysis. (A) show mortality by feeding 1000 ng/µL of <i>BC-Hsp70</i> and <i>BC-CLIC</i> dsRNA over time. Statistical differences between treatments at the same time points are shown by different letters (P<0.05). (B) Shows mortality induced by decreasing concentrations (1000ng/µL, 500 ng/µL, 100ng/µL) of <i>BC-Actin</i> or <i>BC-ATPase</i> dsRNAs. Statistical differences in mortality rates were evaluated between test dsRNAs and GFP dsRNA at the same concentrations and time points. Single asterisk indicates P<0.05, and double asterisks indicate P<0.01. Statistical differences are also shown by different letters between different concentrations for the same dsRNA at the same time points. In Figure (A) and (B) the same data are used for GFP 1000.</p>", "links"=>[], "tags"=>["time-response", "psyllids", "allowed", "diets", "containing"], "article_id"=>382930, "categories"=>["Molecular Biology", "Genetics", "Plant Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Dose_and_time_response_of_psyllids_allowed_to_feed_on_artificial_diets_containing_dsRNAs_/382930", "title"=>"Dose - and time-response of psyllids allowed to feed on artificial diets containing dsRNAs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-16 00:48:50"}
  • {"files"=>["https://ndownloader.figshare.com/files/712677"], "description"=>"<p>The dsRNAs were added to the artificial diet at a concentration of 700 ng/µL. After the teneral adult psyllids were allowed to feed for 72 h, total RNAs were isolated from individual live psyllids by TRIzol method (Invitrogen) and used for cDNA preparation and quantitative real-time PCR. GFP dsRNA-fed served as control for each experiment. (A) show accumulation of <i>BC-ATPase</i> mRNAs in whole psyllid after feeding on dsRNAs for <i>BC-ATPase.</i> (B) show the mRNA abundance of <i>BC-Actin</i> mRNAs in psyllid fed on dsRNAs for <i>BC-Actin</i>. Whole psyllid and dissected guts from a pool of 30 psyllids were analyzed separately. The qRT- PCR results were normalized to the level of <i>B. cockerelli</i> rRNA. Mean values from experimental replicates were analyzed by using Bonferroni (Dunn) <i>t</i>-test to determine between-group significance. Double asterisks indicate differences at the P<0.01 level. The mRNA level in the GFP dsRNA-fed group was arbitarily designated as 1 in all experiments.</p>", "links"=>[], "tags"=>["endogenous", "psyllid", "mrnas", "dsrna"], "article_id"=>383037, "categories"=>["Molecular Biology", "Genetics", "Plant Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Knockdown_of_endogenous_psyllid_mRNAs_by_dsRNA_feeding_/383037", "title"=>"Knockdown of endogenous psyllid mRNAs by dsRNA feeding.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-16 00:50:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/712754"], "description"=>"<p>(A) Small RNAs (21nt) were produced in vitro from GFP and <i>BC-ATPase</i> dsRNAs by digestion with ShorCut RNase III. The small RNAs were separated by 15% acrylamide gel electrophoresis and stained with ethidium bromide. The arrowhead indicates the position of 21nt siRNAs. The sizes of markers are indicated to the right. (B) show qRT-PCR results for <i>BC-ATPase</i> mRNAs after feeding on 100 ng/µL <i>BC-ATPase</i> siRNA for 72 h. GFP siRNA-fed psyllids served as control for treatment. The qRT-PCR results were normalized to the level of <i>B. cockerelli</i> rRNA. Differences between control GFP siRNA-treated samples and <i>BC-ATPase</i> siRNA-treated groups were calculated and shown as P values using the Bonferroni (Dunn) t-test. Double asterisks indicate p<0.01. The mRNA level in the dsGFP group was arbitarily designated as 1 in the experiment.</p>", "links"=>[], "tags"=>["endogenous", "psyllid", "mrnas", "sirna"], "article_id"=>383113, "categories"=>["Molecular Biology", "Genetics", "Plant Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Knockdown_of_endogenous_psyllid_BC_ATPase_mRNAs_by_siRNA_feeding_/383113", "title"=>"Knockdown of endogenous psyllid <i>BC-ATPase</i> mRNAs by siRNA feeding.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-16 00:51:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/712863"], "description"=>"<p>Teneral adult psyllids were allowed to feed for 72h on artificial diet containing 700 ng/µL of <i>BC-Actin</i>, <i>BC-ATPase</i> or GFP dsRNAs. Subsequently, small RNAs were isolated from group of ∼80 individuals, and 1 µg small RNA was separated on a 15% PAGE gel containing 8M Urea and transferred to a nylon membrane. <sup>32</sup>P-UTP-labeled negative strand <i>BC-Actin</i> (A), <i>BC-ATPase</i> (B) or GFP (C) RNA transcripts were used as a probe for the corresponding blot. MicroRNA markers were analyzed on the same gel and sizes are indicated to the right of each blot. As a positive control, a <i>TMV</i> vector pJL36 containing <i>BC-Actin, BC-ATPase</i> or GFP fragment was agro-infiltrated into <i>N. benthamiana</i> and siRNAs were isolated after 2 weeks. (A) Lane 1, pJL36-<i>BC-Actin</i>-infected <i>N. benthamiana</i>; Lane 2, <i>BC-Actin</i> dsRNA-fed psyllids; Lane 3, GFP dsRNA-fed psyllids; Lane 4, sucrose-fed psyllids. The arrow on the left indicates the position of the siRNAs. (B) Lane 1, pJL36-<i>BC-ATPase</i>-infected <i>N. benthamiana</i>; Lane 2, <i>BC-ATPase</i> dsRNA-fed psyllids; Lane 3, GFP dsRNA-fed psyllids; Lane 4, sucrose-fed psyllids. (C) Lane 1, pJL36-GFP-infected <i>N. benthamiana</i>; Lane 2, GFP dsRNA-fed psyllids; Lane 3, GFP dsRNA-fed psyllids were transferred to and fed on tomato seedling for 48h; Lane 4, GFP dsRNA-fed psyllids were transferred to and fed on tomato seedling for 96h; Lane 5, sucrose-fed psyllids. The panels at the bottom show ethidium bromide stained rRNA as a loading control. The exposure times for <i>BC-Actin</i>, <i>BC-ATPase</i> and GFP were 15 h, 19 h and 8 h, respectively.</p>", "links"=>[], "tags"=>["blot", "detection", "rna", "dsrna-fed"], "article_id"=>383227, "categories"=>["Molecular Biology", "Genetics", "Plant Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Northern_blot_detection_of_small_RNA_in_BC_Actin_dsRNA_fed_psyllids_/383227", "title"=>"Northern blot detection of small RNA in <i>BC-Actin</i> dsRNA-fed psyllids.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-16 00:53:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/712943"], "description"=>"<p>1. Each individual was injected with 200 nL of 100 ng/µL dsRNA.</p><p>2. About half of the injected psyllids would die on day 1 after injection and this is most likely caused by the injection procedure.</p><p>3. The percentages of surviving individuals on day 2 - 6 were based on the NO. of surviving individuals on day 1 for each case.</p>", "links"=>[], "tags"=>["induced", "injection", "gfp"], "article_id"=>383305, "categories"=>["Molecular Biology", "Genetics", "Plant Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.t001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mortality_induced_by_injection_of_BC_Actin_and_GFP_dsRNAs_/383305", "title"=>"Mortality induced by injection of <i>BC-Actin</i> and GFP dsRNAs.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-11-16 00:55:05"}
  • {"files"=>["https://ndownloader.figshare.com/files/712985"], "description"=>"<p>1. Psyllids were fed with dsRNA or siRNAs as indicated.</p><p>2. Total RNA was isolated from individual psyllid or gut samples from a pool of 30 insects. WS, whole insect.</p><p>3. The number of psyllids used for qRT-PCR analysis in one experiment. For each experiment, the same numbers of GFP samples were used as controls for test sample. <sup>+</sup>For gut sample, a pool of 30 psyllids was used for every treatment.</p><p>4. The name of sequences used for dsRNA feeding and mRNA detection by qRT-PCR.</p><p>5. The mRNA abundance of specific genes after introduction of corresponding dsRNA or SiRNA was shown as test sample, and the average value of the control GFP group was designated as 1. Expression of each mRNA was normalized to the level of rRNA in the same sample.</p><p>6. Difference between GFP group and test group was calculated and shown as P value using the Bonferroni (Dunn) t-test. Single asterisk indicates p<0.05 and double asterisk indicates p<0.01.</p>", "links"=>[], "tags"=>["Real-time", "pcr", "detection", "endogenous", "mrnas", "feeding", "dsrnas"], "article_id"=>383341, "categories"=>["Molecular Biology", "Genetics", "Plant Biology"], "users"=>["Hada Wuriyanghan", "Cristina Rosa", "Bryce W. Falk"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027736.t002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Quantitative_real_time_PCR_detection_for_endogenous_B_cockerelli_mRNAs_after_feeding_of_dsRNAs_or_siRNAs_/383341", "title"=>"Quantitative real-time PCR detection for endogenous <i>B. cockerelli</i> mRNAs after feeding of dsRNAs or siRNAs.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-11-16 00:55:41"}

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