**2.1 Morphological diversities among 4 species of** *Dendrolimus*

For investigating the morphological diversity, many pupae of *D. punctatus* Walker, *D. superans* Butler, *D. houi* Lajonquiere and *D. kikuchii* Matsumura were collected in 2006(table1).


Table 1. Origin of 4 different species of the tested *Dendrolimus* materials

Genetic Diversity and Population Differentiation of Main Species

of *Dendrolimus* (Lepidoptera) in China and Influence of Environmental Factors on Them 323

YJM

ZJKL

GYYN

LPS

Fig. 1. UPGMA cluster based on morphological traits of 4 different species of the tested

*Dendrolimus* materials

Morphological diversities among 4 species of *Dendrolimus* by analyzing characters such as pupae weight, pupae length, female weight, female wing length, female length, male weight, male wing length and male length. Analysis of variance for all characteristics was significantly different among species and among individuals within populations. Coefficient of variation of morphological traits showed that pupae weight is significantly higher than other traits. Pupae length, female weight and female wing length were the main traits account for the phenotypic variations according to the PCA and discriminate analysis. Based on the UPGMA cluster, four species of *Dendrolimus* may be divided into two groups: the first branch in the dendrogram included *D. punctatus* Walker and *D. superans* Butler, and *D. houi* Lajonquiere and *D. kikuchii* Matsumura were grouped together into second branch(table2-3,figure1)


Note:PW:pupae weight, PL: pupae length, FW: female weight, FWL: female wing length, FL: female length, MW: male weight, MWL: male wing length, ML: male length.

Table 2. Coefficient of variation of morphological traits of 4 different species of the tested *Dendrolimu* materials


Note: PW: pupae weight, PL: pupae length, FW: female weight, FWL: female wing length, FL: female length, MW: male weight, MWL: male wing length, ML: male length.

Table 3. Eigenvalue and principle proportion of 8 character of 4 different species of the tested *Dendrolimus* materials

#### **2.2 Genetic diversities among 4 species of** *Dendrolimus* **by allozyme**

Gene diversity and genetic structure among 4 species of *Dendrolimus* were assessed by allozyme, which provides a powerful tool for investigation of genetic variation (table4,figure2).

A total of 12 presumed loci and 22 alleles were scored in the sampled populations by allozyme method, which 6 loci were polymorphic. At population level, the mean number of alleles per locus (*A*) was 1.2708, the percentage age of polymorphic loci (*P*) was 25.00℅, and the mean expected heterozygosity per locus (*He*) was 0.0828. At species level, *A*=1.8333,

Morphological diversities among 4 species of *Dendrolimus* by analyzing characters such as pupae weight, pupae length, female weight, female wing length, female length, male weight, male wing length and male length. Analysis of variance for all characteristics was significantly different among species and among individuals within populations. Coefficient of variation of morphological traits showed that pupae weight is significantly higher than other traits. Pupae length, female weight and female wing length were the main traits account for the phenotypic variations according to the PCA and discriminate analysis. Based on the UPGMA cluster, four species of *Dendrolimus* may be divided into two groups: the first branch in the dendrogram included *D. punctatus* Walker and *D. superans* Butler, and *D. houi* Lajonquiere and *D. kikuchii* Matsumura were grouped together into second

Code (PW) (PL) (FW) (ML) (FL) (MW) (MWL) (FWL) Mean YJM 54.15 14.13 20.15 4.38 6.01 27.30 4.53 7.87 17.32 GYYN 46.62 12.89 12.85 5.20 3.72 23.09 3.53 2.59 13.81 LPS 58.14 5.21 26.56 8.29 6.00 32.13 4.65 7.58 18.57 ZJKL 49.04 9.68 33.52 4.03 8.57 50.87 9.15 9.21 21.76 Total 51.99 10.48 23.27 5.48 6.08 33.35 5.47 6.81 17.87

Note:PW:pupae weight, PL: pupae length, FW: female weight, FWL: female wing length,

PW 6.0824 76.03 76.03 PL 1.0501 13.13 89.16 FW 0.3807 4.76 93.92 FWL 0.2334 2.92 96.83 FL 0.0886 1.11 97.94 MW 0.0734 0.92 98.86 MWL 0.0515 0.64 99.50 ML 0.0400 0.50 100.00

Table 2. Coefficient of variation of morphological traits of 4 different species of the tested

code Eigenvalue Contribution rate % Cumulative contribution rate %

Note: PW: pupae weight, PL: pupae length, FW: female weight, FWL: female wing length, FL: female

Gene diversity and genetic structure among 4 species of *Dendrolimus* were assessed by allozyme, which provides a powerful tool for investigation of genetic variation

A total of 12 presumed loci and 22 alleles were scored in the sampled populations by allozyme method, which 6 loci were polymorphic. At population level, the mean number of alleles per locus (*A*) was 1.2708, the percentage age of polymorphic loci (*P*) was 25.00℅, and the mean expected heterozygosity per locus (*He*) was 0.0828. At species level, *A*=1.8333,

Table 3. Eigenvalue and principle proportion of 8 character of 4 different species of the

**2.2 Genetic diversities among 4 species of** *Dendrolimus* **by allozyme** 

FL: female length, MW: male weight, MWL: male wing length, ML: male length.

length, MW: male weight, MWL: male wing length, ML: male length.

branch(table2-3,figure1)

*Dendrolimu* materials

tested *Dendrolimus* materials

(table4,figure2).

Fig. 1. UPGMA cluster based on morphological traits of 4 different species of the tested *Dendrolimus* materials

Genetic Diversity and Population Differentiation of Main Species

information index;

*Dendrolimus* materials

tested *Dendrolimus* materials

of pine caterpillars(figure9).

materials

of *Dendrolimus* (Lepidoptera) in China and Influence of Environmental Factors on Them 325

population *A Ae* I *P(%) Ho He* YJM 1.3333 1.1462 0.1403 33.33 0.0944 0.0921 ZJKL 1.1667 1.1569 0.1129 16.67 0.1458 0.0828 GYYN 1.3333 1.1325 0.1207 25.00 0.0877 0.0771 LPS 1.2500 1.1424 0.1258 25.00 0.0972 0.0856 Mean 1.2708 1.1445 0.1249 25.00 0.1063 0.0828 Overall loci 1.8333 1.5564 0.3795 50.00 0.1076 0.2323

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

Ho: Observed heterozygosity; He: Expected heterozygosity; P: Percentage of polyporphic loci Table 5. Genetic diversity parameters of 4 different species of the tested *Dendrolimus* 

Note: Nei's genetic identity (above diagonal) and genetic distance (below diagonal).

**2.3 Genetic diversities among 3 species of** *Dendrolimus* **by ISSR** 

population(*D. spectabilis*), which is identical with Nei′s index(table8-10).

Table 6. Nei's genetic identity and genetic distance of 4 different species of the tested

Fig. 3. UPGMA dendrograms based on Nei's genetic distance of 4 different species of the

A set of optimized response system of ISSR has been developed, which provides a powerful tool for investigation of genetic variation of 9 natural populations(*D. punctatus spectabilis*; *D. punctatus tabulaeformis*; *D. superans*) (figure4-8,table7). Shannon index shows highest genetic diversity in Weichang population(*D. superans*) and lowest genetic diversity in Shenyang

Different geographical groups in the same populations has a genetic divergence in some extent. Genetic similarities and cluster analysis shows that inter-species heredity difference is obviously bigger than the homogeneous between populations in the heredity difference, which reflects the heredity difference degree is consistent from the DNA level with the phenotype level. *D. punctatus spectabilis* has nearer genetic distance with *D. punctatus tabulaeformis* than *D. superans*, which is identical with that researched in sexual information

population YJM ZJKL GYYN LPS YJM \*\*\*\* 0.9142 0.8319 0.7157 ZJKL 0.0897 \*\*\*\* 0.7845 0.6747 GYYN 0.1840 0.2427 \*\*\*\* 0.7967 LPS 0.3345 0.3935 0.2272 \*\*\*\*


Table 4. The kinds of allozymes be choosed

Fig. 2. Electrophoretic forms of LDH, ADH, FDH, GDH, MDH, ME From left to right: SYC, QDC, JZC, HTM, TDM, YJM

*P*= 50.00℅, *He* = 0.2323. So, the genetic diversity at species level was rather high than that at population level (table5,6). A UPGMA dendrogram was generated based on *Nei's* genetic distance showed 4 species of *Dendrolimus* were clustered into two groups: the first branch in the dendrogram included *D. punctatus* Walker and *D. superans* Butler, and the second branch included *D. houi* Lajonquiere and *D. kikuchii* Matsumura (figure3).

LDH MDH ME ADH FDH GDH CAT POD

E.C. 1. 1. 1. 27 E.C. 1. 1. 1. 37 E.C. 1. 1. 1. 40 E.C. 1. 1. 1. 1 E.C. 1.2. 1. 2 E.C. 1.4. 1.2 E.C. 1. 11. 1. 6 E.C. 1. 1. 1. 1

*Fdh*-1

*Me-*1

*Me-*2

*Mdh-*1

*Mdh-*2

Enzyme Abbreviation E.C. code

*Adh*-1

*Gdh-*1

*P*= 50.00℅, *He* = 0.2323. So, the genetic diversity at species level was rather high than that at population level (table5,6). A UPGMA dendrogram was generated based on *Nei's* genetic distance showed 4 species of *Dendrolimus* were clustered into two groups: the first branch in the dendrogram included *D. punctatus* Walker and *D. superans* Butler, and the second branch

Fig. 2. Electrophoretic forms of LDH, ADH, FDH, GDH, MDH, ME

included *D. houi* Lajonquiere and *D. kikuchii* Matsumura (figure3).

From left to right: SYC, QDC, JZC, HTM, TDM, YJM

1. LDH 2. MDH

*Ldh-*2

*Ldh*-1

*Ldh*-3

4. FDH

6. ME

Lactate dehydrogenase Malate dehydrogenase

Alcohol dehydrogenase Formate dehydrogenase Glutamate dehydrogenas

Malic enzyme

Catalase Peroxidase

3. ADH

5. GDH

Table 4. The kinds of allozymes be choosed


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 5. Genetic diversity parameters of 4 different species of the tested *Dendrolimus*  materials


Note: Nei's genetic identity (above diagonal) and genetic distance (below diagonal).

Table 6. Nei's genetic identity and genetic distance of 4 different species of the tested *Dendrolimus* materials

Fig. 3. UPGMA dendrograms based on Nei's genetic distance of 4 different species of the tested *Dendrolimus* materials
