Insect systematics+species interaction
Chao-Dong Zhu
zhucd@ioz.ac.cn*Key Laboratory of Zoological Systematics and Evolution, Institute of Zoology,
Chinese Academy of Sciences*College of Biological Sciences, University of Chinese Academy of Sciences

08.07.2021

Outline

Institute of Zoology, Chinese Academy of Sciences

1. Insect Systematics

2. Species Interactions

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Functional Insect Groups

Pollinators
Predators
Parasitoids
Herbivores
Decomposers

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Pollinator Insects

玉米螟

大豆蚜虫朱砂叶螨（棉红蜘蛛）

褐飞虱Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Parasitoid Wasps

玉米螟大豆蚜虫

朱砂叶螨
（棉红蜘蛛）

褐飞虱Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Scientific Questions

Prof. Robert May in Oxford：‘‘How many distinct life forms – species does your planet have?’’（May 1986, 1992, 1993, Nature; May 1988, 2010, Science）

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Morphology-based / 2003
Taxonomy

DNA Taxonomy

Characters

Sequences

Insect Taxonomy

Insect Phylogeny

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Morphology-based/ 2003
Major Sampling Methods

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Morphology-based/ 2003

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Morphology-based/ 2003

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Morphology-based/ 2003
In China, we catalogued more than 1372 known bee
species, published more than 70 papers after 2009, withmany more unknown yet to sciences.

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Morphology-based/ 2003
There are more than 1691 researchersand students joined us on insect
identification via QQ. They are working
on insect taxonomy, ecology, plant
protection, quarantine and pollination
biology.

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Morphology-based/ 2003
Morphology based taxonomists have been in decline intorare specialists. However, more demands for accurate
identifications from agriculture, ecology. How can we fill in
the gap?

 With a lot of unkown species, we need to speed up
taxonomy; With very large molecular data, we need to provide
taxonomy supports

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006
Taxonomy

DNA Taxonomy

Characters

SequencesInsect Taxonomy

Insect Phylogeny

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006
1. Quick and Efficient DNA based identification system

0.2AF-003B
Outgroup
AF-015
A4-013B C. bivittatus
A3-056A C. ceroplastae
A3-045A C. ceroplastae
A3-054A C. ceroplastae
A3-054B C. ceroplastae
A2-004A C. ceroplastae
A3-022E C. ceroplastae
A2-006A C. ceroplastae
A2-006C C. ceroplastae
A1-025A C. ceroplastaeA1-025E C. ceroplastae
A3-016G C. ceroplastae
A3-016F C. ceroplastae
A3-052A C. ceroplastae
A2-014A C. ceroplastae
A3-016E C. ceroplastae
A5-118C C. ceroplastes
A1-015B C. ceroplastae
A4-003A C. ceroplastae
A1-012A C. ceroplastae
A1-015H C. ceroplastae
A4-003C C. ceroplastae
A4-006B C. ceroplastae
A4-006A C. ceroplastae
A4-006C C. ceroplastae
A3-043I C. ceroplastae
A1-019E C. sp5
A3-021A C. sp5
A1-019I C. sp5
A3-032A C. sp5
A1-019C C. sp5
A1-019H C. lycimnia
A1-019A C. lycimnia
A1-006B C. lycimnia
A3-017 C. lycimnia
A2-021B C. lycimnia
A3-020B C. lycimnia
A1-006A C. lycimnia
A3-020A C. lycimnia
A4-001A C. spN
A1-011S C. spN
A4-001E C. spN
A4-001D C. spN
A4-008A C. spXJ
A4-008D C. spXJ
A1-017C C. sp2
A1-017B C. sp2
A1-005A C. sp2
A3-028B C. sp2
E3-186A C. spY
A1-035A C. spY
A1-014A C. spY
A1-014C C. spY
A4-109A C. spF
A4-018A C. nigricorpus
A5-018A C. nigricorpus
A1-045C C. yoshidae
A1-045A C. yoshidae
A09-01A C. yoshidae
A09-01C C. yoshidae
A1-011D C. yoshidae
A4-014C C. yoshidae
A3-030A C. semicircularis
A3-023D C. semicircularis
A1-027D C. ishiii
A1-011A C. ishiii
A3-013A C. ishiii
A1-027B C. ishiii
A3-050A C. ishiii
A4-002A C. ishiii
A3-057B C. pseudococci nr
A3-061A C. aterrimus
A5-004A C. bogoriensis
A3-038A C. bogoriensis
A3-040H C. bogoriensis
A3-040A C. bogoriensis
A3-040D C. bogoriensis
A4-011A C. japonicus
A1-002 C. japonicus
A4-016B C. japonicus
A1-046C C. japonicus
A1-011C C. japonicus
A1-011U C. japonicus
A1-011V C. japonicus
A2-003E C. japonicus
A2-003C C. japonicus
A2-003F C. japonicus
A3-036A C. japonicus
A3-036B C. japonicus
A1-007 C. japonicus
A3-055A C. japonicus
A5-117C C. japonicus
A1-041A C. japonicus
A4-011C C. japonicus
A11-5 C. japonicusA3-044C C. japonicus
A3-011A C. japonicus
A5-032B C. japonicus
A4-005B C. japonicus
A1-037A C. japonicus
A4-005A C. japonicus
A1-029 C. japonicus
A5-033A C. japonicus
A11-1 C. japonicus
A1-036A C. japonicus
A1-020B C. japonicus
A4-003D C. japonicus
A3-051A C. japonicus
A3-044A C. japonicus
A2-009D C. japonicus
A3-051D C. japonicus
A2-009G C. japonicus
A2-009A C. japonicus
A2-009B C. japonicus
A3-014C C. japonicus
A1-020C C. japonicus
A1-010 C. japonicus
A1-016D C. japonicus

ABGD
NJ

GMYC
COI
CombineBayesian
NJ
BayesianPTP

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006
2. Single Locus Species Delimitation（Hebert 2003, Proc. R. Soc. Lond. B）

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006
All PNGs have similar efficiency to distinguish species（Luo …, Cameron*, Zhu* 2011, BMC Genomics）

Insect Systematics

Institute of Zoology, Chinese Academy of SciencesDNA Taxonomy / 2006

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006
3. Multiple Loci Taxonomy（Chesters…, Zhu*, 2015, Methods in Ecology and Evolution）

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006



3 gene dataset for 250 bee specimens
Novel heuristic search of clustering parameters
Figure shows 70 global species units

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006
4. Species delimitation in HUGE databaseAbout 80000 insect species wereautomaticallyand quickly identified based on 24gene loci

（Chesters and Zhu*, 2015, Systematic Biology)

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006
5. Large insect phylogenies

Chesters, 2017, Systematic Biology

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

DNA Taxonomy / 2006
6. Species delimitation across a range ofspeciation scenarios

Luo…, Zhu*, 2018, Systematic Biology

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

NGS Taxonomy / 2016
Developing a new pipeline

High-quality Assemble Genome
Detecting species with very low abundanceMore accurately on abundance

Zhang*…, Zhu, 2019, Genome Biology & Evolution

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

NGS Taxonomy / 2016

Insect Systematics

Institute of Zoology, Chinese Academy of Sciences

Total-evidence Taxonomy / 2021
SiteSpecies
1 1 0 1 1 1 0 0 10 1 0 1 1 0 0 0 10 1 0 1 1 1 1 0 10 0 0 0 1 1 1 0 0

Goodwin et al., 2016

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Scientific Questions

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Scientific Questions

Producer

Consumer(herbivore,pollinator)

Predator/parasitiod

Symbiotic-bacteria

Interaction of multi-trophic levels
Gradient of tree diversity

Biodiversity?
Network?
Co-existence?
Stability?
……

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

BEF-China Field Site

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

DNA Approaches
Morphology or DNA?
An example from plantcaterpillars-parasitoids

Fig. 4. Quantitative tri-trophic food web for secondary rainforest
vegetation in PNG comprising 37 plant species, 4803 caterpillars
from 154 species (exposed and semi-concealed guilds) and 643
parasitoids from 76 species (mostly Braconidae, Tachinidae andIchneumonidae).

Hrcek et al. 2013,
Oecologia

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

DNA Approaches
Monitoring biodiversity via metabarcoding

Ji et al. Ecology Letters, 2013

More bioinformatics
required for large dataanalyses

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

DNA Approaches
Overview of NGS-based molecular methods for biodiversity assessment.

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

DNA Approaches
Building on previous work on bee
diversity in and around rubber
plantationsPollen obtained from common bees
such as the eastern honey bee
Plant library constructed for localspecies
2 pollen barcodes sequenced using
Illumina platform

Liu et al. (2017). Scientific Reports.

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 1. Nest Trap System• Limited but representative
number of species（pollinator、predator、parasitoid）• Cheap, standardized, readily
available• Quantifying trophic interactions（pollen - bee，prey – wasp，
host - parasitoid）• Assessment of species’ entire lifehistory

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 1. Nest Trap System
Adult broke theseal
cleptoparasitism
Larva broke thesealparasitism
Monoparasitoid
parasitoid
Hyperparasitoid

Melittobia sp.

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 1. Nest Trap SystemYear
2015-2016
Number oftubes
5321
Number ofcells
13790
Number ofparasitoids
1598

2018
2312
5164
493

Tree diversity gradientsparasitic rate

Hymenoptera hostTrap nest

morphospecies
parasitoid

Hymenoptera hostspecies richness andabundanceParasitoid species richness
and abundanceSpecies Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 1. Nest Trap System
The ecological effect of tree diversity on cavity-nesting Hymenoptera andassociated parasitoids Abundance and species richness of
predatory wasps and parasitoids werepositively correlated with tree species
richness, while bee abundance andbee species richness were unrelated
to tree species richness.
 tree species richness increases theabundance and species richness ofimportant communities such aspredators and parasitoids.

Guo,…Li, … Zhu*. 2020. Basic and applied Ecology

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 2. Predator System
The ecological effect of tree diversity on predator (spider) diversity.

Natural enemy hypothesis: Predators

tend to be polyphagous and have broad habitat
requirements, so they would be expected toencounter a greater array of alternative preyand microhabitats in a heterogeneousenvironment
Most related studies were conducted infarmland and grassland, not forest.

Q1：How do predators’ species diversity respond to tree diversity？
Q2：How do predators’ functional traits respond to tree diversity？

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 2. Predator System

AdultsImmaturity
Sampling20172018

20192020

Total4449

26851707

2751

April

1862

961

385

NAJune

1577

782

370

973

September

1010

942

952

1779

Identification

Reference databaseof spiders

Sequencing

Sequencing

measuring

species diversity
phylogenetic diversity
Function diversity

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 2. Predator System Abundance and species
richness of spiders werepositively correlated withtree functional richness

Chen,…Zhu. In preparing

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 3. Herbivore SystemYear20172018

Sampling in 2017-2018
Rearing & Sequencing
BeatingMonth
Number of larvae
April(7-24)
718
June(7-7.6)
2053
September(1-15)
472
April(7-20)
June(6-17)
September( 3-15 )Barcodelibrary 1

Larvae

1162
3488
574

Moth Adults

Barcodelibrary 2

LarvaeParasitoids Adults

Identification & Sequencing
Light trap

Moth Adults

Barcodelibrary 3

Identifiedspecies

Species Interactions

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Project 3. Herbivore System
3.1 The ecological effect of tree diversity on herbivore diversity

 Lepidopteran abundance, the effective number of species and Faith's PD and MPD ,but not MNTD,
significantly increased with experimentally manipulated tree species richness.

Wang, Li, … Zhu*. 2019. Journal of Ecology

Species Interactions

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Project 3. Herbivore System
3.2 Host functional and phylogenetic composition structure plant–herbivore networks
 phylogenetic host composition andrelated palatability/defence traits
but not tree species richness
significantly affected herbivore
communities and interaction
network complexity at both thespecies and community levels

Wang, Li, … Zhu*. 2020. Molecular Ecology

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 3. Herbivore System
3.3 Tree species richness and leaf traits shaped/driven caterpillar-associated microbiomes

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 3. Herbivore System
3.3 Tree species richness and leaf traits shaped/driven caterpillar-associated microbiomes

 7,909 bacterial OTUs from 634
caterpillar individuals
 Different caterpillar species
feeding on the same tree

species showing the similar
pattern of microbial
composition.

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 3. Herbivore System
3.3 Tree species richness and leaf traits shaped/driven caterpillar-associated microbiomes
 Tree diversity and leaf traits (Leaf toughness anddry matter content) was found to drive the diversityof caterpillar-associated bacteria both directly andindirectly via effects on caterpillar communities

Li,…,Zhu*. 2020. Ecology and Evolution

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 3. Herbivore System
3.3 Tree species richness and leaf traits shaped/driven caterpillar-associated microbiomes

 Microbial α-diversity: soil > leaf
litter > leaves > caterpillar

 Phyllosphere is one of the main

sources of the herbivore microbiome

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 3. Herbivore System
3.4 Phylogenetic relatedness, functional traits and spatial scale determine herbivore
co-occurrence Lepidoptera co-occurrence was negatively associated with
their phylogenetic relatedness, tree diversity and defensive
traits, but positively associated with nutritional functionaltraits. Further, species co-occurrence was higher at larger
spatial scales (tree species or plot) than at smaller scale(individual trees).

Wang, Li, … Zhu*. in revision

Species Interactions

Institute of Zoology, Chinese Academy of SciencesProject 4. Functional Traits

Specimenµ-Computer Tomography (µ-CT)
3D Reconstruction

Critical Point Dryer
Scanning Electron Microscope (SEM)

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 4. Functional Traits
Transfer descriptive morphology into digital bioinformatic pipeline:(1) Undamaged anatomy and more detailed characters(2) Phylogenetic and geometric morphometrics(3) Functional morphological data for ecological comparison and modeling(4) Finite element analysis for forces and material properties

Species Interactions

Institute of Zoology, Chinese Academy of Sciences

Project 4. Functional Traits
Question: What kinds of morphological characters
assist the larvae have such a powerful mouthpartcrossing phylogenetic barriers?
Experiment:(1) Carefully examine the eating processes ofphylogenetically multiple lepidopteran larvae(2) Make 3D reconstruction for their mouthparts(3) Referring to the corresponding plantmorphological characters, summarize theimportant mouthpart characters of larvae

Mouthpart of larvae

Additional experiment:
3D reconstruction of mouthpart of special feeding
habits (e.g. leaf mining)
Labial palp

Acknowledgements

Institute of Zoology, Chinese Academy of Sciences

Team MembersAndreas Schuldt

Ming-Qiang Wang

Institute of Zoology, Chinese Academy of Sciences

Douglas Chesters

Qing-Song Zhou

Si-Pei Liu

Yi LiPeng-Fei Guo

Jing-Ting Chen