野莴苣及其近似种间亲缘关系研究

林玲; 虞赟; 陈智明; 王德玲; 沈建国; 伏建国

doi:10.19303/j.issn.1008-0384.2017.10.006

野莴苣及其近似种间亲缘关系研究

doi: 10.19303/j.issn.1008-0384.2017.10.006

林玲^{1, 2,},
虞赟^2, ,,
陈智明³,
王德玲⁴,
沈建国²,
伏建国⁵

1.
泉州出入境检验检疫局, 福建泉州 362000
2.
福建出入境检验检疫局技术中心, 福建福州 350001
3.
福州出入境检验检疫局, 福建福州 350015
4.
福建农林大学, 福建福州 350002
5.
江苏出入境检验检疫局, 江苏南京 210000

基金项目:

国家质检总局科技计划项目 2015IK043

国家质检公益性行业科研专项 201410076

福建省科技计划重大专项 2017NZ0003-1-7

详细信息

作者简介:
林玲(1980-), 女, 农艺师, 研究方向:植物检疫(E-mail:303880739@qq.com)

通讯作者:
虞赟(1981-), 女, 高级农艺师, 研究方向:植物检疫(E-mail:3900091@qq.com)

中图分类号: Q78
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- 被引次数: 0
出版历程
- 收稿日期: 2017-07-16
- 修回日期: 2017-09-08
- 刊出日期: 2017-10-28

Genetic Comparisons Between Lactuca pulchella (Pursh) DC and Its Closely Related Lettuce Species

LIN Ling^{1, 2
,},
YU Yun^{2
, ,},
CHEN Zhi-ming³,
WANG De-ling⁴,
Shen Jian-guo²,
Fu Jian-guo⁵

1.
Quanzhou Entry-Exit Inspection and Quarantine Bureau, Quanzhou, Fujian 362000, China
2.
Inspection & Quarantine Technology Center, Fujian Entry-Exit Inspection and Quarantine Bureau/Fujian Key laboratory for Technology Research of Inspection and Quarantine, Fuzhou, Fujian 350001, China
3.
Fuzhou Entry-Exit Inspection and Quarantine Bureau, Fuzhou, Fujian 350015, China
4.
Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China
5.
Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing Jiangsu 210000, China

摘要

摘要: 利用PCR方法对野莴苣及其11种近似种DNA进行扩增，获得ITS、matK 和rbcL 序列，分析遗传相似系数和聚类分析图谱，探讨野莴苣及其近似种的亲缘关系。通过野莴苣及其近似种ITS序列分析发现：野莴苣和生菜、莴苣、毒莴苣的相似性分别为93.6%、93.1%、92.6%，野莴苣与乳苣的相似性仅为87.8%；matK 序列研究发现：野莴苣和生菜、莴苣、毒莴苣的相似性分别为99.2%、99.0%、99.1%，而野莴苣与乳苣的相似性仅为98.3%，结果表明，野莴苣与莴苣属的生菜、莴苣、毒莴苣的亲缘关系更近，与乳苣属的乳苣亲缘关系次之。通过对比rbcL 序列发现：野莴苣和生菜相似度达100%，与莴苣、翅果菊、毒莴苣、刺毛莴苣、宿根莴苣的相似度均达99.7%，而野莴苣与乳苣的相似度却为99.4%，将野莴苣归为莴苣属更加合理。
- 野莴苣 /
- 近似种 /
- 亲缘关系
Abstract: The genetic similarity coefficients and cluster analysis maps on Lactuca pulchella (Lp) and its 11 closely related species were obtained by using PCR to amplify ITS, matK and rbcL gene from the total DNA fragments of the lettuce varieties. Based on the ITS sequences, a similarity between Lp and L. sativa var. ramose was found to be 93.6%; between Lp and L. sativa , 93.1%; between Lp and L. serriola , 92.6%;and, between Lp and Mulgedium tataricum , the lowest at 87.8%. According to the matK sequences, a similarity between Lp and L. sativa var. ramose was shown to be 99.2%;between Lp and L. sativa , 99.0%, ; between Lp and L. serriol , 99.1%; and, between Lp and M.tataricum , 98.3%. It appeared that Lp was related closer to the Lactuca species than Mulgedium . By comparing the rbcL , Lp and L. sativa L. var. ramosa had a perfect match with a 100% similarity; whereas, Lp and L. sativa , L. indica , L.serriola , L. hirsute or L. perennis , at 99.7%;and, Lp and M. tataricum , at 99.4%. Consequently, it was concluded that Lp belonged to the genus Lactuca .
- Lactuca pulchella /
- closely related species /
- genetic relationship

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图 1 野莴苣及其近似种的聚类分析

Figure 1. Cluster analysis on L. pulchella and its close relative species

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表 1 供试材料名称

Table 1. Test materials

试验编号	名称	拉丁名
A1	生菜	Lactuca sativa L. var. ramosa Hort.
A2	苣荬菜	Sonchus arvensis L.
A3	莴苣	L.sativa L.
A4	苦苣菜	S. oleraceus L.
A5	翅果菊	L. indica L.
A6	毒莴苣	L. serriola L.
A7	刺毛莴苣	L. hirsuta Muhl. ex Nutt.
A8	黄鹌菜	Youngia japonica (L.)DC.
A9	乳苣	Mulgedium tataricum (L.) DC.
A10	宿根莴苣	L. perennis L.
A11	雀苣	L. viminea L.
A12	野莴苣	L.pulchella (Pursh) DC.

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表 2 PCR所用引物序列

Table 2. Primers used for PCR

引物名称	引物序列(5'-3')	目标片断 /bp
ITS-F	GGAAGTAAAAGTCGTAACAAG	722
ITS-R	TCCTCCGCTTATTGATATGC
matK -F	ATACCTTACCCAGCCCATCT	580
matK -R	AGCATTTGACTGCGTACCAT
rbcL -F	ATGTCACCACAAACAGAAAC	718
rbcL -R	TCGCATGTACCTGCAGTAGC

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表 3 PCR扩增反应体系

Table 3. PCR amplification reaction system

成分	体积/μ L
2×Taq PCR MasterMix	25
引物-F	2
引物-R	2
ddH₂O	18
DNA template	3
总体积	50

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表 4 野莴苣及其近似种ITS序列的遗传相似系数

Table 4. Genetic similarity coefficients of L. pulchella and its closely related species based on ITS sequences

(单位/%)
样品编号	A1	A2	A3	A4	A5	A6	A7	A8	A9	A10	A11	A12
A1	100
A2	81.0	100
A3	100	82.5	100
A4	80.3	91.2	81.9	100
A5	91.2	81.4	91.8	81.6	100
A6	100	82.8	100	82.1	91.9	100
A7	88.7	77.2	88.7	78.0	96.3	88.7	100
A8	82.6	80.6	83.9	80.4	82.1	84.1	77.2	100
A9	93.0	81.7	93.5	80.8	91.5	93.6	87.8	83.5	100
A10	85.7	78.4	85.4	77.1	87.0	85.6	86.1	78.0	85.0	100
A11	74.6	74.8	75.7	73.9	75.9	75.8	71.1	74.6	76.7	74.5	100
A12	93.6	78.4	93.1	77.3	87.3	92.6	84.6	80.4	87.8	82.5	74.2	100
注：A1为生菜；A2为苣荬菜；A3为莴苣；A4为苦苣菜；A5为翅果菊；A6为毒莴苣；A7为刺毛莴苣；A8为黄鹌菜；A9为乳苣；A10为宿根莴苣；A11为雀苣；A12为野莴苣。下表同。

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表 5 野莴苣及其近似种matK 序列的遗传相似系数

Table 5. Genetic similarity coefficients of L. pulchella and its closely related species based on matK sequences

(单位/%)
样品编号	A1	A2	A3	A4	A5	A6	A7	A8	A9	A10	A11	A12
A1	100
A2	97.9	100
A3	99.8	98.0	100
A4	97.7	99.2	97.7	100
A5	98.1	96.7	98.6	96.9	100
A6	99.8	98.0	100	97.7	98.6	100
A7	98.6	97.5	99.0	97.1	97.9	98.9	100
A8	98.1	98.2	98.2	97.9	97.3	98.2	97.3	100
A9	98.1	96.7	98.6	96.9	100	98.6	97.9	97.3	100
A10	98.8	97.3	98.8	97.1	97.7	98.9	98.1	97.8	97.7	100
A11	96.7	97.3	97.4	96.7	96.7	97.3	97.0	97.6	96.7	96.5	100
A12	99.2	97.5	99.0	97.5	98.2	99.1	98.4	97.8	98.3	98.1	96.7	100

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表 6 野莴苣及其近似种rbcL 序列的遗传相似系数

Table 6. Genetic similarity coefficients of L. pulchella and its closely related species based on rbcL sequences

(单位/%)
样品编号	A1	A2	A3	A4	A5	A6	A7	A8	A9	A10	A11	A12
A1	100
A2	99.1	100
A3	100	99.0	100
A4	99.0	100	99.1	100
A5	100	99.0	100	99.1	100
A6	100	99.0	100	99.1	100	100
A7	100	99.0	100	99.1	100	100	100
A8	98.9	99.2	99.0	99.2	99.0	99.0	99.0	100
A9	99.8	98.7	99.7	98.7	99.7	99.7	99.7	98.7	100
A10	100	93.0	93.6	99.1	93.9	94.0	93.6	93.5	93.7	100
A11	99.2	99.2	99.2	99.5	99.2	99.2	99.2	99.3	98.9	92.0	100
A12	100	98.8	99.7	99.1	99.7	99.7	99.7	98.6	99.4	99.7	98.9	100

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参考文献(18)

[1]	中华人民共和国进境植物检疫性有害生物名录(更新至2017年6月, 441种)[OL]. [2017-06-05]. http://www.aqsiq.gov.cn/xxgk_13386/zvfg/gfxwj/dzwjy/201706/t20170614_490858.htm
[2]	Flora of North America Editorial Committee. Flora of North America, North of Mexico[M]. New York:Oxford University Press, 2006, 19-21:259.
[3]	Flora of Missouri Editorial Committee.Flora of Missouri[OL].[2017-06-05].http://www.efloras.org/florataxon.aspx?flora_id=11&taxon_id=200024120.
[4]	United States Department of Agriculture. Natural Resources Conservation Service.[OL].[2017-06-05]. http://plants.usda.gov/core/profile?symbol=LATAP.
[5]	中国科学院中国植物志编辑委员会.中国植物志[M].北京:科学出版社, 1997, 80:75.
[6]	韩亚光.新侵入辽宁地区的杂草——野莴苣[J].沈阳农业大学学报, 1995, 26(1):77-79. http://www.cqvip.com/qk/93761X/199501/index.shtml
[7]	梁照文, 童明龙, 高振峰, 等.毒莴苣及其近似种的形态比较[J].植物检疫, 2014, 28(4):36-37. http://www.cqvip.com/QK/93407X/201404/661724391.html
[8]	Flora of China Editorial Committee.Flora of China (Asteraceae)[M]. Beijing & St. Louis:Science Press & Missouri Botanical Garden Press, 2011, 20-21:234, 237, 238.
[9]	KRESS W J, WURDACK K J, ZIMMER E A, et al. Use of DNA barcodes to identify flowering plants.[J]. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102(23):8369-8374. http://www.doc88.com/p-086650407796.html
[10]	TABERLET P, COISSAC E, POMPANON F, et al. Power and limitations of the chloroplast trnL (UAA) intron for plant DNA barcoding[J]. Nucleic Acids Research, 2007, 35(3):e14. doi: 10.1093/nar/gkl938
[11]	KRESS W J, ERICKSON D L. A two-locus global DNA barcode for land plants:the coding rbcL gene complements the non-coding trnH-psbA spacer region[J]. Plos One, 2007, 2(6):e508. doi: 10.1371/journal.pone.0000508
[12]	FAZEKAS A J, BRUGESS K S, KESANAKURTI P R, et al. Multiple Multilocus DNA Barcodes from the Plastid Genome Discriminate Plant Species Equally Well[J]. Plos One, 2007, 3(7):e2802. http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.275.1938
[13]	LAHAYE R, VAN D B M, BOGARIN D, et al. DNA barcoding the floras of biodiversity hotspots[J]. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105(8):2923-2928. doi: 10.1073/pnas.0709936105
[14]	伏建国, 杨晓军, 钱路, 等.植物DNA条形码技术在出入境检验检疫领域的应用[J].植物检疫, 2012, 26(2):64-69. http://www.cqvip.com/qk/93407X/201202
[15]	付涛, 王志龙, 钱萍仙, 等.高等植物DNA条形码最新研究进展及其应用[J].核农学报, 2016, 30(5):887-896. doi: 10.11869/j.issn.100-8551.2016.05.0887
[16]	FU Y M, JIANG W M, FU C X. Identification of species within Tetrastigma (Miq.) Planch.(Vitaceae) based on DNA barcoding techniques[J].植物分类学报, 2011, 49(3):237-245. http://www.cnki.com.cn/Article/CJFDTOTAL-ZWFX201103010.htm
[17]	LI D Z, GAO L M, LI H T, et al. Comparative analysis of a large dataset indicates that internal transcribed spacer (ITS) should be incorporated into the core barcode for seed plants.[J]. Proceedings of the National Academy of Sciences, 2011, 108(49):19641-19646. doi: 10.1073/pnas.1104551108
[18]	高连明, 刘杰, 蔡杰, 等.关于植物DNA条形码研究技术规范[J].植物分类与资源学报, 2012, 34(6):592-60. http://ir.kib.ac.cn/handle/151853/16324