**2.7 Genetic diversities among 5 sub-populations of** *D. punctatus tabulaeformis* **Tsai** *et* **liu**

The pupae of 5 sub-populations of *D. punctatus tabulaeformis* were collected in Pingquan, Hebei Province(table28). And the gene diversity and genetic structure of them were assessed by SSR (table29).


Table 28. Origin of the tested *D. punctatus tabulaeformis* materials in Chengde City, Pingquan county


Genetic Diversity and Population Differentiation of Main Species

M: molecular marker(PBR322∕Msp I marker) Fig. 16. SSR profile amplified by primer SSR5

M: molecular marker(PBR322∕Msp I marker) Fig. 17. SSR profile amplified by primer SSR6

527 bp

110 bp

527 bp

110 bp

of *Dendrolimus* (Lepidoptera) in China and Influence of Environmental Factors on Them 339

1~6, 7~12, 13~18, 19~24, 25~30 are pop1, pop2, pop3, pop4, pop5 of *D. punctatus tabulaeformis*;

1~5,6~10,11~15,16~20,21~25 are pop1, pop2, pop3, pop4, pop5 of *D. punctatus tabulaeformis*;

three populations were grouped together into second branch (table31,figure18).

gene flow among sub-populations. A UPGMA dendrogram based on *Nei's* genetic distance showed 5 sub-populations of *D. punctatus tabulaeformis* Tsai *et* liu were clustered into two groups: the first branch in the dendrogram included pop1 and pop2, and the remaining


Table 29. SSR primers of the tested *D. punctatus tabulaeformis* materials in this study

For genetic diversities, among 5 sub-populations of *D. punctatus tabulaeformis*, in total, polymorphic bands were produced using 8 primers in SSR analysis method (table29, figure15-17). The percentage of polymorphic loci (*P*) was 80.00%, the mean number of alleles per locus (*A*) was 2.6250, and the mean expected heterozygosity per locus (*He*) was 0.3765. For the species level, *P*=87.50%, *A*=3.1250, *He*=0.4747(table30). From the summary of Fstatistics at polymorphic loci of 6 populations, we could found the *Fst* was 0.2159, which mean there show high genetic diversity among populations.The number of migrants per generation among populations (*Nm*) was 0.9081, suggesting the occurrence of rather low

1~5,6~10,11~15,16~20,21~25 are pop1, pop2, pop3, pop4, pop5 of *D. punctatus tabulaeformis*; M: molecular marker (PBR322Msp I marker)

Fig. 15. SSR profile amplified by primer SSR4

1~6, 7~12, 13~18, 19~24, 25~30 are pop1, pop2, pop3, pop4, pop5 of *D. punctatus tabulaeformis*; M: molecular marker(PBR322∕Msp I marker)

Fig. 16. SSR profile amplified by primer SSR5

338 The Dynamical Processes of Biodiversity – Case Studies of Evolution and Spatial Distribution

Table 29. SSR primers of the tested *D. punctatus tabulaeformis* materials in this study

1~5,6~10,11~15,16~20,21~25 are pop1, pop2, pop3, pop4, pop5 of *D. punctatus tabulaeformis*;

For genetic diversities, among 5 sub-populations of *D. punctatus tabulaeformis*, in total, polymorphic bands were produced using 8 primers in SSR analysis method (table29, figure15-17). The percentage of polymorphic loci (*P*) was 80.00%, the mean number of alleles per locus (*A*) was 2.6250, and the mean expected heterozygosity per locus (*He*) was 0.3765. For the species level, *P*=87.50%, *A*=3.1250, *He*=0.4747(table30). From the summary of Fstatistics at polymorphic loci of 6 populations, we could found the *Fst* was 0.2159, which mean there show high genetic diversity among populations.The number of migrants per generation among populations (*Nm*) was 0.9081, suggesting the occurrence of rather low

allele

3

1

2

3

4

3

3

6

Size range (bp)

78~188

161

155~160

217~333

193~207

234~430

187~220

158~241

Tm(°C)

52

50

50

50

51

52

52

50

Primer Sequences (5'—3') Number of

TCATCCCGAGTCCCACTCA ATTGCTCTTCCTATCTGGCTA GTTCTCGGTCGTGGTTTTAG AACCGCTTCCGCCGATTAC CTGGCACCCCGCCGATTAC AACAAAACAATTATAAACTCTTAC ACCAACTTCGACACCTTCT CACTGCCCCGAACCTATAC ACTTCTACTGCGTGTGAACT GTCCCTTTGTCCGATAATATG GGAGCACCAATGAAGAATGT GTTTCTACCTCATGGGATCTTTTAGCTC ACGTAAAACTAATCAA CTGTCCAAAGCAAACTATC CTGCTAGAGCTTTCTGTGTT AAGAATTTCAATTTAAGACTGAC

SSR4

SSR5

SSR6

SSR7

SSR8

SSR9

SSR10

SSR11

527 bp

110 bp

M: molecular marker (PBR322Msp I marker) Fig. 15. SSR profile amplified by primer SSR4

1~5,6~10,11~15,16~20,21~25 are pop1, pop2, pop3, pop4, pop5 of *D. punctatus tabulaeformis*; M: molecular marker(PBR322∕Msp I marker)

Fig. 17. SSR profile amplified by primer SSR6

gene flow among sub-populations. A UPGMA dendrogram based on *Nei's* genetic distance showed 5 sub-populations of *D. punctatus tabulaeformis* Tsai *et* liu were clustered into two groups: the first branch in the dendrogram included pop1 and pop2, and the remaining three populations were grouped together into second branch (table31,figure18).

Genetic Diversity and Population Differentiation of Main Species

*Dendrolimus*

*tabulaeformis* **populations** 

Community Tree

Community Sample

I

II

III

matter and nitrogen in soil(table32-36).

so main factor could not be judged(table37).

height

Note: I,II: pure forests; III: mixed forest.

plot

Note: I,II: pure forests; III: mixed forest

Diameter at breast height

III 8.5 13.5 0.5 Half

Table 32. Statistical results of stand condition investigation

Species number

of *Dendrolimus* (Lepidoptera) in China and Influence of Environmental Factors on Them 341

**3.1 The effect of environmental conditions on the genetic diversity of** *D. punctatus* 

The genetic diversity among 3 natural populations of the Chinese pine caterpillar (*Dendrolimus punctatus tabulaeformis*) were tested by AFLP method in one *Pinus tabulaeformis*-*Dahurian larch* mixed forest and two *Pinus tabulaeformis* pure forests in Pingquan county. Besides, investigations on plant species diversity, forest crown density, incidence extent, gradient and exposure of three forest communities were taken, while dilution heat method and semi-micro Macro Kjeldahl method were used to determine the content of organic

The result of principle component analysis shows that the growth status is main factor which influent genetic diversity of *D. punctatus tabulaeformis* populations. Besides, site conditions had some effects on it. An integrated effect was produced by all site conditions,

Mixed forest has a great influence on gene flow among different populations of *D. punctatus tabulaeformis*, because gene flow between populations in mixed forest and in pure forests was lower than that of two pure forests. Gene flow between populations in pure forest with larger species abundance and in mixed forest was higher than between population in pure forest with lower species abundance and in mixed forest, which showed that the correlation between gene flow among different populations and species abundance of pine forests is negative.

Crown

I 7.9 11.6 0.6 Ubac 31.7° 1 II 7.8 10.9 0.5 Adret 34.3° 2

> Individual number

Table 33. Species diversity index analysis of 3 populations in *D. punctatus tabulaeformis*

density Exposure Gradient Incidence

Simpson s diversity index

1 20 285 0.7962 2.9628 2 16 327 0.8400 2.9936 3 17 307 0.8619 3.1770

4 25 499 0.8722 3.3170 5 15 728 0.8387 2.9772 6 20 392 0.8184 2.9746

7 29 1021 0.8363 3.3473 8 31 868 0.8627 3.4626 9 29 949 0.8055 3.1790

adret 36.0° 3

extent

Shannon-weaver diversity index

**3. The effect of environmental conditions on the genetic diversity of** 


Note: *A:* Number of alleles per locus; *Ae:* Effective number of alleles per locus; *I:* The Shannon information index;

*Ho:* Observed heterozygosity; *He:* Expected heterozygosity; *P:* Percentage of polyporphic loci

Table 30. Genetic diversity parameters of 5 sub-populations in *D. punctatus tabulaeformis*


Note: See table 3-1 for abbreviations of population codes.

Table 31. Nei's genetic identity and genetic distance of 5 sub-populations in *D. punctatus tabulaeformis*

Fig. 18. Phylogenetic relationship of 5 sub-populations of *D. punctatus tabulaeformis* based on Nei's distance of SSR markers and clustered using UPGMA
