生物多样性 ›› 2013, Vol. 21 ›› Issue (3): 315-325.doi: 10.3724/SP.J.1003.2013.11035

• • 上一篇    下一篇

家蚕地方品种遗传多样性及其分子系统学研究

杜周和1, 2, 刘俊凤1, *(), 刘彬斌1, 左艳春1, 吴建梅1, 陈义安1, 张剑飞1, 鲁成2, *()   

  1. 1 四川省农科院蚕业研究所, 四川南充 637000
    2 西南大学蚕学与系统生物学研究所, 重庆 400716
  • 收稿日期:2013-02-04 接受日期:2013-04-26 出版日期:2013-05-20
  • 通讯作者: 刘俊凤,鲁成 E-mail:liujunfeng1993@263.net;lucheng@swu.edu.cn
  • 基金项目:
    国家重点基础研究发展计划(2005CB121000)、四川省应用基础(2013JY0113)和省农科院优秀论文基金(2013LWJJ-008)

Genetic diversity and molecular genealogy of local silkworm varieties

Zhouhe Du1, 2, Junfeng Liu1, *, Binbin Liu1, Yanchun Zuo1, Jianmei Wu1, Yi-an Chen1, Jianfei Zhang1, Cheng Lu2, *()   

  1. 1 Institute of Sericulture, Sichuan Academy of Agricultural Sciences, Nanchong, Sichuan 637000
    2 Institute of Sericulture and Systems Biology, Southwest University, Chongqing 400716
  • Received:2013-02-04 Accepted:2013-04-26 Online:2013-05-20
  • Contact: Liu Junfeng,Lu Cheng E-mail:liujunfeng1993@263.net;lucheng@swu.edu.cn

为检测家蚕(Bombyx mori)地方品种间的遗传差异, 分析其遗传多样性和系统分化模式, 为家蚕的品种鉴定和杂交育种亲本选择提供分子依据, 本文对72个家蚕地方品种的淀粉酶基因(Bmamy2)片段进行测序, 采用DNA序列分析技术探究了家蚕地方品种在核苷酸序列水平上的遗传差异及其系统分化特征。结果显示, 家蚕地方品种在Bmamy2基因变异丰富, 碱基突变率为5.6-8.2%, 单倍型多样度为0.8294, 核苷酸多样度为0.0236±0.00122, 表明Bmamy2是一个具有丰富序列多样性的标记基因, 能够较好地区分家蚕地方品种间的遗传多样性。系统发育分析、分子方差分析和Network分析等多种分析方法均检测到家蚕地方品种不按地理系统或生态类型聚类, 各聚类种群均由来自不同地理系统或生态类型的品种构成, 而来自同一地理系统或生态类型的品种则分别归属于不同的聚类种群。推测在家蚕的进化过程中并没有形成按地理系统或生态类型归类的分化格局, 家蚕可能具有多种化性混合驯化起源的遗传背景和系统分化特征。

关键词: 家蚕, 种质资源, 遗传多样性, 分子系统学

In order to explore the genetic diversity and systematic differentiation pattern among silkworm varieties, aiming to guide hybridization breeding, we sequenced a total of 72 Bmamy2gene fragments from local silkworm varieties. The analysis of nucleotide sequence diversity and systematic differentiation indicated that there was rich genovariation in the sequencing region of Bmamy2gene, and the base mutation rate is 5.6- 8.2%, the haplotype diversity is 0.8294, and the nucleotide diversity is 0.0236±0.00122, suggesting Bmamy2being a better marking gene with rich nucleotide sequence diversity, based on which the genetic diversity among different local silkworm varieties can be identified. The same heredity population structure is proclaimed by several analysis methods that every clade consisting of varieties from different geosystems and ecological types, while the varieties from the same geosystem and ecotype belong to different clades in the phylogeny. There is no population structure pattern that different varieties claded together according to geosystem or ecotype. It can be speculated that the silkworm origins from mixture of kinds of several voltinism mulberry silkworm, Bombyx mandarina, while the domestication events took place in several regions, from which the domesticated mulberry silkworms are all devoting to the domesticated silkworm population of today.

Key words: Bombyx mori, germplasm resources, genetic diversity, molecule genealogy

表1

供试的72个家蚕品种材料的来源及特性"

序号
No.
品种
Variety
代号
Code
生态类型
Ecotype
序号
No.
品种
Variety
代号
Code
生态类型
Ecotype
1 小石丸
Xiaoshiwan
XSW CV1 37 萨尼斯
Sanisi
SNS EV1
2 二毛
Ermao
EM CV1 38 罗尼7
Luoni7
LN7 EV1
3 土三虎斑
Tusanhuban
TSHB CV1 39 法408
Fa408
F408 EV1
4 中土
Zhongtu
ZT CV1 40 欧18
Ou18
O18 EV1
5 达县土种
Daxiantuzhong
DXTZ CV1 41 意16
Yi16
Y16 EV1
6 土一肉黄圆
Tuyirouhuangyuan
TYRHY CV1 42 乌D
WuD
WD EV2
7 土八
Tuba
Tba CV1 43 乌G
WuG
WG EV2
8 鲁1
Lu1
L1 CV1 44 乌B
WuB
WB EV2
9 Re黑
Rehei
RH CV1 45 乌2
Wu2
W2 EV2
10 鲁10
Lu10
L10 CV1 46 日110
Ri110
R110 T
11 土种01
Tuzhong01
TZ01 CV1 47 大造
Dazao
DZ T
12 诸桂
Zhugui
ZG CV1 48 高花
Gaohua
GH T
13 温州种
Wenzhouzhong
WZZ CV1 49 第五白卵
Diwubailuan
DWBL T
14 芙蓉灰卵
Furonghuiluan
FRHL CV1 50 演金黄白
Yanjinhuangbai
YJHB T
15 灰色卵
Huiseluan
HSL CV1 51 15-100 15-100 T
16 盱眙种
Xuyizhong
XYZ CV1 52 07-310 07-310 T
17 余杭24
Yuhang24
YH24 CV1 53 高白
Gaobai
GB T
18 沛县种
Peixianzhong
PXZ CV1 54 琼山海南
Qiongshanhainan
QSHN T
19 邯郸种
Handanzhong
HDZ CV1 55 655 655 T
20 中4010
Zhong4010
Z4010 CV2 56 防四
Fangsi
FS T
21
Tai
T CV2 57 印度种
Yinduzhong
YDZ T
22 玉溪2号
Yuxi2hao
YX CV2 58 加秋
Jiaqiu
JQ T
23
Ping
P CV2 59 白夏B
BaixiaB
BXB T
24 C108 C108 CV2 60 天龙青白
Tianlongqingbai
TLQB JV1
25 余杭2
Yuhang2
YH2 CV2 61 赤熟
Chishu
Cshu JV1
26 土白
Tubai
Tbai CV2 62 日9
Ri9
R9 JV1
27 松花形吴
Songhuaxingwu
SHXW CV2 63 姬蚕
Jican
JC JV1
28 保黄
Baohuang
BH CVd 64 黑子
Heizi
HZ JV1
29 MCH101 MCH101 CVd 65 日10
Ri10
R10 JV1
30 BT924 BT924 CVd 66 绵蚕
Miancan
MC JV1
31 BH863 BH863 CVd 67 日限2
Rixian2
RX2 JV2
表1 (续) Table 1 (continued)
序号
No.
品种
Variety
代号
Code
生态类型
Ecotype
序号
No.
品种
Variety
代号
Code
生态类型
Ecotype
32 琼十
Qiongshi
QS CVd 68 赢纹形
Yingwenxing
YWX JV2
33 四海
Sihai
SH CVd 69 J115 J115 JV2
34 兰十
Lanshi
LanS CVd 70 830 830 JV2
35 罗萨
Luosa
LuoS EV1 71 春四
Chunsi
CS JV2
36 阿利可斯
Alikesi
ALKS EV1 72 T8 T8 JV2

图1

家蚕Bmamy2基因结构(F、R分别代表上下游引物)"

图2

Bmamy2基因扩增区序列结构(黑框代表外显子, 斜线框代表内含子)"

表2

72个家蚕地方品种Bmamy2基因扩增片段碱基变化统计"

序列长度
Sequence length (bp)
碱基插入/缺失位点数
Insert/delete (%)
变异位点数
Variable site (%)
单个突变位点
Singleton variable site (%)
简约信息位点
Parsimony variable site (%)
外显子 Exon 493 2(0.4)* 10(2.0) 2(0.4) 8(1.6)
内含子 Intron 670 139(20.7) 55(8.2) 7(1.0) 48(7.2)
全序列 Entire 1,163 141(12.1) 65(5.6) 9(0.8) 56(4.8)

表3

不同生态类型家蚕品种间的遗传距离"

CV2 EV1 T CV1M3 JV2 CV1M4 JV1 CVd
CV2
EV1 0.025
T 0.027 0.024
CV1M3 0.026 0.015 0.023
JV2 0.026 0.012 0.024 0.016
CV1M4 0.026 0.013 0.024 0.017 0.014
JV1 0.027 0.025 0.024 0.025 0.026 0.025
CVd 0.025 0.025 0.024 0.025 0.026 0.025 0.024
EV2 0.025 0.044 0.029 0.039 0.044 0.042 0.026 0.026

图3

家蚕地方品种单倍型网络图(H代表单倍型, mv代表可能存在的单倍型, 生态类型代号见表1)"

图4

基于Bmamy2基因核苷酸序列的家蚕NJ树(MEGA4.0建树, 模型选择: P-距离, 成对删除gaps, 1,000次重复, 节点处仅显示>40%的bootstrap值)。品种代号见表1。"

表4

各聚类支家蚕品种构成"

聚类支
Clade
地理生态类型
Ecotype
品种数(个)
No. of varieties
占各支百分比
%
A 中系一化 CV1 15 36.59
中系二化 CV2 4 9.76
中系多化 CVd 3 7.32
日系一化 JV1 3 7.32
日系二化 JV2 5 12.20
欧系一化 EV1 6 14.63
热带系统 T 5 12.20
小计 Sum 41
B 中系一化 CV1 6 33.33
中系多化 CVd 2 11.11
日系一化 JV1 2 11.11
日系二化 JV2 1 5.56
欧系一化 EV1 1 5.46
热带系统 T 6 33.33
小计 Sum 18
C 中系二化 CV2 4 30.77
中系多化 CVd 2 15.38
日系一化 JV1 2 14.38
欧系二化 EV2 4 30.77
热带系统 T 1 7.69
小计 Sum 13
合计 Total 72

表5

家蚕72个品种Bmamy2基因的分子变异(按地理生态类型分类)"

变异来源
Source of variance
自由度
df
方差分量
Variance component
变异百分比
(%)
P-value
地理生态类型组群间 Among geo-voltings 7 6.93028 19.86 > 0.05
地理生态类型组群内 Among population within geo-voltings 1 1.23267 3.53 > 0.05
种群内 Within populations 63 26.73978 76.61 < 0.01
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