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Usually a large number of samples are required to identify insect diets in food web studies. Previously, Sanger sequencing and next generation sequencing (NGS) with short DNA barcodes were used, resulting in low species\u2010level identification; meanwhile the costs of Sanger sequencing are expensive for metabarcoding together with more samples. Here, we present a fast and effective sequencing strategy to identify larvae of Lepidoptera and their diets at the same time without increasing the cost on Illumina platform\u00a0in a single HiSeq run, with long\u2010multiplex\u2010metabarcoding (<jats:italic>COI<\/jats:italic> for insects, <jats:italic>rbcL<\/jats:italic>, <jats:italic>matK<\/jats:italic>, <jats:italic>ITS<\/jats:italic> and <jats:italic>trnL<\/jats:italic> for plants) obtained by Trinity assembly (SHMMT). Meanwhile, Sanger sequencing (for single individuals) and NGS (for polyphagous) were used to verify the reliability of the SHMMT approach.\u00a0Furthermore, we show that SHMMT approach is fast and reliable, with most high\u2010quality sequences of five DNA barcodes of 63 larvae individuals (54 species) recovered (full length of 100% of the <jats:italic>COI<\/jats:italic> gene and 98.3% of plant DNA barcodes) using Trinity assembly (up\u2010sized to 1015 bp). For larvae diets identification, 95% are reliable; the other 5% failed because their guts were empty. The diets identified by SHMMT approach are 100% consistent with the host plants that the larvae were feeding on during our collection. Our study demonstrates that SHMMT approach is reliable and cost\u2010effective for insect\u2010plants network studies. This will facilitate insect\u2010host plant studies that generally contain a huge number of samples.<\/jats:p>","DOI":"10.1111\/1744-7917.12749","type":"journal-article","created":{"date-parts":[[2019,12,27]],"date-time":"2019-12-27T14:50:51Z","timestamp":1577458251000},"page":"127-143","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Plant\u2013herbivorous insect networks: who is eating what revealed by long barcodes using high\u2010throughput sequencing and Trinity assembly"],"prefix":"10.1111","volume":"28","author":[{"given":"Xiao\u2010Man","family":"Zhang","sequence":"first","affiliation":[{"name":"College of Life Sciences Capital Normal University  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhi\u2010Yong","family":"Shi","sequence":"additional","affiliation":[{"name":"College of Life Sciences Capital Normal University  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shao\u2010Qian","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Life Sciences Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peng","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Life Sciences Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John\u2010James","family":"Wilson","sequence":"additional","affiliation":[{"name":"Vertebrate Zoology at World Museum National Museums Liverpool  Liverpool United Kingdom"},{"name":"Department of Microbiology and Parasitology, Faculty of Medical Science Naresuan University  Phitsanulok Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chungkun","family":"Shih","sequence":"additional","affiliation":[{"name":"College of Life Sciences Capital Normal University  Beijing China"},{"name":"Department of Paleobiology National Museum of Natural History, Smithsonian Institution  Washington DC USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Li","sequence":"additional","affiliation":[{"name":"College of Life Sciences Capital Normal University  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xue\u2010Dong","family":"Li","sequence":"additional","affiliation":[{"name":"College of Life Sciences Capital Normal University  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guo\u2010Yue","family":"Yu","sequence":"additional","affiliation":[{"name":"Institute of Plant and Environment Protection Beijing Academy of Agriculture and Forestry Sciences  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3450-5421","authenticated-orcid":false,"given":"Ai\u2010Bing","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Life Sciences Capital Normal University  Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,4,13]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2891-9-23"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-294X.2009.04412.x"},{"key":"e_1_2_10_4_1","first-page":"52","volume-title":"Fauna Sinica Insecta (Vol.16) Lepidoptera: Noctuidae","author":"Chen Y.X.","year":"1999"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1093\/aesa\/88.4.590"},{"key":"e_1_2_10_6_1","first-page":"8","article-title":"Introducing the bipartite package: analysing ecological networks","volume":"8","author":"Dormann C.F.","year":"2008","journal-title":"R News"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/1755-0998.12429"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.en.34.010189.000313"},{"key":"e_1_2_10_9_1","first-page":"1","article-title":"Life histories of Indian insects","volume":"6","author":"Fletcher T.B.","year":"1920","journal-title":"Microlepidoptera. 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