生物多样性 ›› 2017, Vol. 25 ›› Issue (6): 627-637.doi: 10.17520/biods.2017090

• 研究报告 • 上一篇    下一篇

结合分子手段和形态分析验证大白杜鹃与马缨杜鹃的自然杂交

郑硕理1, 2, 田晓玲3, 黄承玲3, 王灵军4, 冯元4, 张敬丽1, *()   

  1. 1 云南农业大学园林园艺学院, 昆明 650201
    2 湖南省森林植物园, 长沙 410116
    3 贵州民族大学人文科技学院, 贵阳 550025
    4 贵州百里杜鹃科研所, 贵州毕节 551518
  • 收稿日期:2017-03-22 接受日期:2017-06-28 出版日期:2017-06-20
  • 通讯作者: 张敬丽 E-mail:jl200812@yeah.net
  • 基金项目:
    国家自然科学基金(31260079)和国家科技支撑计划(2015BAD10B00)

Molecular and morphological evidence for natural hybridization between Rhododendron decorum and R. delavayi (Ericaceae)

Shuoli Zheng1, 2, Xiaoling Tian3, Chengling Huang3, Lingjun Wang4, Yuan Feng4, Jingli Zhang1, *()   

  1. 1 College of Landscape and Gardening, Yunnan Agriculture University, Kunming 650201
    2 Hunan Forest Botanical Garden, Changsha 410116
    3 College of Humanities & Science, Guizhou Minzu University, Guiyang 550025
    4 Institute of Guizhou Baili Rhododendron, Bijie, Guizhou 551518
  • Received:2017-03-22 Accepted:2017-06-28 Online:2017-06-20
  • Contact: Zhang Jingli E-mail:jl200812@yeah.net

中国西南地区是世界杜鹃属(Rhododendron)植物的分布和分化中心, 开展杜鹃属自然杂交研究可为分类修订和新品种选育提供依据。本文以贵州百里杜鹃自然保护区(百里保护区)和云南石宝山分布的大白杜鹃(Rhododendron decorum)、马缨杜鹃(R. delavayi)及两种间疑似自然杂交后代和人工杂交后代个体为研究材料, 通过形态特征比较、核基因间隔序列ITS以及叶绿体基因片段trnL-trnF、rbcL和trnH-psbA测序特征分析, 发现疑似自然杂交后代与人工杂交后代形态特征介于大白杜鹃与马缨杜鹃之间; 4个DNA片段序列均能区分大白杜鹃和马缨杜鹃, 同一物种的不同居群在叶绿体基因片段rbcL、trnH-psbA序列存在差异位点; 所有人工杂交后代个体和1个石宝山疑似杂交个体均以大白杜鹃为母本, 所有百里保护区疑似杂交个体和6个石宝山疑似杂交个体以马缨杜鹃为母本。本研究证实了大白杜鹃与马缨杜鹃在百里保护区和云南石宝山存在自然杂交现象, 杂交为双向且自然杂交后代绝大多数以马缨杜鹃为母本。

关键词: 杜鹃属, 自然杂交, 形态特征, DNA序列

The southwestern region of China is the center of differentiation for the genus Rhododendron. Studies on the natural hybridization of Rhododendron provide solutions for taxonomic problems and enhance the ability to breed new cultivars faster. In this paper, we compared the morphological features and sequenced nuclear internal transcribed spacer (ITS) and chloroplast trnL-trnF, rbcL, trnH-psbA for Rhododendron decorum, R. delavayi, and the putative hybrid offspring of these two species. All samples in this study were collected from Baili Rhododendron Nature Reserve in Guizhou Province, the Shibao Mountain of Yunnan Province and Kunming Botanical Garden. Results showed that all putative hybrids (natural and artificial) produced an intermediate morphology. Sequencing results revealed that all artificial hybrids and one collected from the Shibao Mountain inherited cpDNA from R. decorum, and all putative hybrids collected from Baili Rhododendron Nature Reserve and six from Shibao Mountain inherited cpDNA from R. delavayi. In this study, we have confirmed that natural hybridization between R. decorum and R. delavayi exists in the Guizhou and Yunnan sites. Hybridization was bilateral, and most of the hybrids were derived from maternal parent R. delavayi.

Key words: Rhododendron, natural hybridization, morphological traits, DNA sequences

表1

大白杜鹃、马缨杜鹃及疑似杂交后代的来源和数量"

编号 Code 种类 Taxon 地点 Site 经纬度
Latitude and longitude
海拔
Altitude (m)
个体数量
Sample size
BLM (1-10) 马缨杜鹃 R. delavayi 贵州百里杜鹃自然保护区 Baili Rhododendron
Nature Reserve in Guizhou Province
27°15′ N 105°58′ E 1,700-1,800 10
BLD (1-10) 大白杜鹃 R. decorum 贵州百里杜鹃自然保护区 Baili Rhododendron
Nature Reserve in Guizhou Province
27°15′ N 105°58′ E 1,700-1,800 10
BLH (1-8) 疑似杂交后代
Putative hybrids
贵州百里杜鹃自然保护区 Baili Rhododendron
Nature Reserve in Guizhou Province
27°15′ N 105°58′ E 1,700-1,800 8
SBSM (1-5) 马缨杜鹃 R. delavayi 云南大理石宝山 Shibao Mountain of Dali, Yunnan 26°22′ N 99°50′ E 2,400-2,600 5
SBSD (1-5) 大白杜鹃 R. decorum 云南大理石宝山 Shibao Mountain of Dali, Yunnan 26°22′ N 99°50′ E 2,400-2,600 5
SBSH
(2/3/4/5/7/8/9)
疑似杂交后代
Putative hybrids
云南大理石宝山 Shibao Mountain of Dali, Yunnan 26°22′ N 99°50′ E 2,400-2,600 7
AH (1-5) 人工杂交后代
Artificial hybrids
昆明植物园 Kunming Botanical Garden 5
总计 Total 50

表2

大白杜鹃、马缨杜鹃、人工杂交后代与疑似杂交后代的形态特征对比"

形态特征
Morphological characters
大白杜鹃
R. decorum
疑似自然杂交与人工杂交后代
Putative and artificial hybrids
马缨杜鹃
R. delavayi
花冠颜色 Corolla color 白色 White 粉红 Pink 红色 Red
花冠裂片数 No. of corolla lobes 6-8, 通常7 6-8, usually 7 5-6, 无规律 5-6, irregular 5
叶片形状 Leaf shape 长圆形 Oblong 长圆形至披针形 Oblong to lanceolate 披针形 Lanceolate
叶片毛被 Ventral leaf suraface indumentum 无毛 Glabrous 稀疏毛被 Thin indumentum 浓密海绵状毛被
Woolly indumentum
果实表面特征 Fruit surface
character
有腺体 Glands 有腺体或毛被或两者均有
Glands or/and indumentum
密被棕红色毛被 Continuous brownish red indumentum
花期 Flowering period 5-6月 May to June 4月中旬-5月中旬 Mid-April to mid-May 3月-5月 March to May

表3

大白杜鹃与马缨杜鹃的ITS序列差异位点及疑似杂交后代克隆序列"

编号
Code
克隆数量
Clone numbers
类型
Type
差异位点 Variable sites (bp) 类型 Type
94 101 114 203 494 505 A1 A2 A4
大白杜鹃 R. decorum 0 A1 T G G T T C 15 0 0
马缨杜鹃 R. delavayi 0 A2 C C T G C T 0 15 0
AH、BLH、SBSH A3 Y S K K Y Y
克隆测序 Clone sequences
BLH1 2 1 1 0
BLH2 4 1 2 1
BLH3 3 2 1 0
BLH4 3 2 1 0
BLH5 3 1 1 1
BLH6 5 1 1 3
BLH7 4 2 1 1
BLH8 4 2 1 1
AH1 3 1 2 1
AH2 3 1 2 0
AH3 4 2 2 0
AH4 3 2 1 0
AH5 4 2 2 1
SBSH2 6 4 2 0
SBSH3 8 5 3 0
SBSH4 5 4 1 0
SBSH5 5 4 1 0
SBSH7 7 5 2 0
SBSH8 7 6 1 0
SBSH9 5 3 2 0
克隆数量总计
Total clone numbers
53 31 10

表4

大白杜鹃、马缨杜鹃及疑似杂交后代的trnL-trnF序列差异位点"

编号
Code
类型
Type
差异位点 Variable sites (bp)
147 622 623 624 626 627 629 646 831
大白杜鹃 Rhododendron decorum B1 G T T T T T T C T
马缨杜鹃 R. delavayi B2 T A A A A A A A C
AH、SBSH5 B1 G T T T T T T C T
BLH、SHSH2/3/4/7/8/9 B2 T A A A A A A A C

表5

大白杜鹃、马缨杜鹃及疑似杂交后代的rbcL序列差异位点"

编号
Code
类型
Type
差异位点 Variable sites (bp)
8 30 263 398 407
BLD C1 C G T C C
BLM C2 C T A T T
SBSD C3 G G T C C
SBSM C4 G T A T T
AH C1 C G T C C
BLH C2 C T A T T
SBSH5 C3 G G T C C
SBSH2/3/4/7/8/9 C4 G T A T T

表6

大白杜鹃、马缨杜鹃及疑似杂交后代的trnH-psbA序列差异位点"

编号
Code
类型
Type
差异位点 Variable sites (bp)
1 2 128 135 211 214 245 254 254 299 313 333 337 340 344 348 351 358 365-369
BLD D1 G A T C G C G A C A A G T G G T T T AATTC
SBSD D2 - G * * * * * * * * * A A A A A G G *****
BLM D3 * * A G A C T T T T G * * * * * * * G----
SBSM D4 * * A G A A T T T T G * * * * * * * G----
AH D1 G A T C G C G A C A A G T G G T T T AATTC
SBSH5 D2 * G * * * * * * * * * A A A A A G G *****
BLH D3 * * A G A C T T T T G * * * * * * * G----
SBSH2/3/4/7/8/9 D4 * * A G A A T T T T G * * * * * * · G----

图1

大白杜鹃、马缨杜鹃及疑似杂交后代的联合ITS、trnL-trnF、rbcL、psbA-trnH序列片段的聚类图。AH、BLH、SBSH、BLM、BLD、SBSM和SBSD含义见表1。"

图2

大白杜鹃、马缨杜鹃、疑似杂交后代与人工杂交后代的花序和叶片"

图3

大白杜鹃、马缨杜鹃、疑似杂交后代与人工杂交后代部分数量性状。AH、BLH和SBSH含义见表1。"

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