马尾松二代无性系种子园子代父本分析及花粉散布

doi:10.3724/SP.J.1258.2011.00937

摘要/Abstract

摘要：

目前国内较早建立的马尾松(Pinus massoniana)二代种子园正陆续进入正常开花结实期。研究马尾松二代种子园花粉散布和自由授粉子代的父本组成, 可为生产上指导马尾松高世代种子园的规划设计和遗传管理提供理论依据。该文利用筛选的11对SSR引物, 对马尾松二代无性系种子园内8个无性系单株的320个自由授粉子代和48个候选父本进行了扩增, 并采用最大似然法对子代进行父本分析。结果表明: 11个位点共检测到61个等位基因, 每个位点的等位基因数在2-11之间, 平均为5.55个。试验亲本和子代群体的总平均观测杂合度(H_o)、期望杂合度(H_e)及多态信息含量(PIC)分别为0.428、0.433和0.387。在80%的可信度水平下可为232 (72.50%)个子代确定其父本。平均每个采种母树与19个父本产生子代。在自由授粉状态下, 马尾松二代种子园自交率为1.72%, 自交现象很弱, 其交配方式以异交为主。绝大多数亲本无性系的雄性繁殖适合度在1.00%-4.00%之间, 候选父本平均繁殖适合度为2.17%, 平均形成5个后代。马尾松有效花粉的散布距离和固定交配距离的父本繁殖适合度均符合正态分布, 两者呈极显著负相关, 其主要散布距离集中在0-100 m, 而检测到的最大散布距离为192 m。种子园花粉污染率较低, 仅为4.06%。总体看来, 树冠南面子代亲本交配距离较北面有增加的趋势, 但树冠南、北面子代父本组成数并未表现明显的规律。

关键词: 雄性繁殖适合度, 父本分析, 马尾松, 花粉散布, 二代无性系种子园

Abstract:

Aims The earlier second generation seed orchards of Pinus massoniana have been going to normal blossom stage in China. Our objective was to study the pollen dispersal and constitution of the male parents of open-pollination progenies to provide a theoretical basis for the design and genetic management of advanced generation seed orchards.
Methods We identified 320 open-pollination progenies from 8 clone individuals and 48 clones as candidate male parent using 11 polymorphic SSR loci. CERVUS was used to assign the paternity of each progeny based on maximum likelihood analysis.
Important findings We detected 61 alleles at 11 loci, and the number of alleles ranged from 2 to 11 (average of 5.55). The average observed and expected heterozygosity (H_o and H_e) were 0.428 and 0.433, respectively, and the average polymorphic information content (PIC) was 0.387. Among 320 progenies, 232 progenies (75.50%) could be assigned paternity at a confidence level of 80%. Progenies were produced by each mother tree with 19 male trees. The self-crossing rate reached 1.72% in open-pollination condition, and outcrossing was the primary mating mode. The reproduction contribution varied from 1.00 to 4.00% for most male parents, with an average of 2.17% (produced 5 progenies). Strong negative correlation existed between pollination distance and reproductive success of male parents with the same mating distance. In accord with a normal distribution, the main pollination distance ranged from 0 to 100 m; maximum dispersal distance was 192 m. The pollen contamination ratio was 4.06%. Compared with the north progenies of crown, there was an increasing trend of the mating distance of parents in the south progenies of crown, while there was no obvious trend in the male parent numbers of the south and north progenies of crown.

Key words: male reproductive success, paternity analysis, Pinus massoniana, pollen dispersal, second generation clonal seed orchard

谭小梅, 周志春, 金国庆, 张一. 马尾松二代无性系种子园子代父本分析及花粉散布. 植物生态学报, 2011, 35(9): 937-945. DOI: 10.3724/SP.J.1258.2011.00937

TAN Xiao-Mei, ZHOU Zhi-Chun, JIN Guo-Qing, ZHANG Yi. Paternity analysis and pollen dispersal for the second generation clonal seed orchard of Pinus massoniana. Chinese Journal of Plant Ecology, 2011, 35(9): 937-945. DOI: 10.3724/SP.J.1258.2011.00937

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://www.plant-ecology.com/CN/10.3724/SP.J.1258.2011.00937

https://www.plant-ecology.com/CN/Y2011/V35/I9/937

图/表 6

参考文献 31

[1]	Ai C ( 艾畅), Xu LA ( 徐立安), Lai HL ( 赖焕林), Huang MR ( 黄敏仁), Wang ZR ( 王章荣) (2006). Genetic diversity and paternity analysis of a seed orchard in Pinus massoniana. Scientia Silvae Sinicae (林业科学), 42(11), 146-150. (in Chinese with English abstract)
[2]	Botstein D, White RL, Skolnick M, Davis RW (1980). Construction of a genetic linkage map in man using restriction fragment length polymorphisms. The American Journal of Human Genetics, 32, 314-331. URL PMID
[3]	Chaix G, Gerber S, Razafimaharo V, Vigneron P, Verhaegen D, Hamon S (2003). Gene flow estimating with microsatellites in a Malagasy seed orchard of Eucalyptus grandis. Theoretical and Applied Genetics, 107, 705-712. DOI URL PMID
[4]	Chen XY ( 陈小勇), Song YC ( 宋永昌) (2001). Analysis of parentage for naturally established seedlings of Cyclo- balanopsis glauca. Journal of Wuhan Botanical Research (武汉植物学研究), 18(3), 174-180. (in Chinese with English abstract)
[5]	Clark MS (1997). Plant Molecular Biology—A Laboratory Manual. Translated by Gu HY (顾红雅), Qu LJ (瞿礼嘉)(1998). Higher Education Press, Beijing. 3-11. (in Chinese)
[6]	Feng FJ, Sui X, Chen MM, Zhao D, Han SJ, Li MH (2010). Mode of pollen spread in clonal seed orchard of Pinus koraiensis. Journal of Biophysical Chemistry, 1, 33-39.
[7]	Gerber S, Chabrier P, Kremer A (2003). FAMOZ: a software for parentage analysis using dominant, codominant and uniparentally inherited markers. Molecular Ecology Notes, 3, 479-481.
[8]	Goto S, Miyahara F, Ide Y (2002). Identification of the male parents of half-sib progeny from Japanese black pine (Pinus thunbergii Parl.) clonal seed orchard using RAPD markers. Breeding Science, 52, 71-77. DOI URL
[9]	Hansen OK, Kjær ED (2006). Paternity analysis with microsatellites in a Danish Abies nordmanniana clonal seed orchard reveals dysfunctions. Canada Journal of Forest Research, 36, 1054-1058.
[10]	He TH ( 何田华), Ge S ( 葛颂) (2001). Mating system, paternity analysis and gene flow in plant populations. Acta Phytoecologica Sinica (植物生态学报), 25, 144-154. (in Chinese with English abstract)
[11]	Huang SQ ( 黄双全), Guo YH ( 郭友好) (2000). Advances in the study of pollination biology. Chinese Science Bulletin (科学通报), 45, 225-237. (in Chinese)
[12]	Jones ME, Shepherd M, Henry R, Delves A (2008). Pollen flow in Eucalyptus grandis determined by paternity analysis using microsatellite markers. Tree Genetics & Genomes, 4, 37-47.
[13]	Kalinowski ST, Taper ML, Marshall TC (2007). Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Molecular Ecology, 16, 1099-1106. DOI URL PMID
[14]	Lian CL, Miwa M, Hogetsu T (2001). Outcrossing and paternity analysis of Pinus densiflora(Japanese red pine) by microsatellite polymorphism. Heredity, 87, 88-98. URL PMID
[15]	Marshall TC, Slate J, Kruuk LEB, Pemberton JM (1998). Statistical confidence for likelihood-based paternity inference in natural populations. Molecular Ecology, 7, 639-655. DOI URL PMID
[16]	Moriguchi Y, Tani N, Itoo S, Kanehira F, Tanaka K, Yomogida H, Taira H, Tsumura Y (2005). Gene flow and mating system in five Cryptomeria japonica D. Don seed orchards as revealed by analysis of microsatellite markers. Tree Genetics & Genomes, 1, 174-183. DOI URL
[17]	Ritland K, Jain S (1981). A model for the estimation of outcrossing rate and gene frequencies using n independent loci. Heredity, 47, 35-52.
[18]	Robledo-Arnuncio JJ, Gil L (2005). Patterns of pollen dispersal in a small population of Pinus sylvestris L. revealed by total-exclusion paternity analysis. Heredity, 94, 13-22. URL PMID
[19]	Shaw DV, Brown AHD (1982). Optimum number of marker loci for estimating outcrossing in plant populations. Theoretical and Applied Genetics, 61, 321-325. URL PMID
[20]	Shen XH ( 沈熙环) (1994). Seed Orchard Techniques for High Genetic Quality and Ample Production of Seeds (种子园优质高产技术). China Forestry Publishing House, Beijing. 15. (in Chinese)
[21]	Snow AA, Lewis PO (1993). Reproductive traits and male fertility in plants: empirical approaches. Annual Review of Ecology and Systematics, 24, 331-351.
[22]	Sun YG ( 孙亚光), Li HG ( 李火根) (2008). Variations of male reproductive success and sexual selection among individuals of the experimental population of Liriodendron revealed by SSR Markers. Molecular Plant Breeding (分子植物育种), 6(1), 79-84. (in Chinese with English abstract)
[23]	Tan XM ( 谭小梅), Jin GQ ( 金国庆), Zhang Y ( 张一), Qin GF ( 秦国峰), Chu DY ( 储德裕), Zhou ZC ( 周志春) (2011). Genetic variation of flowering and fruiting in the dwarfed second-generation clonal seed orchard of Pinus massoniana. Journal of Northeast Forestry University (东北林业大学学报), 39(4), 39-42. (in Chinese with English abstract)
[24]	Vanden Broeck A, Cottrell J, Quataert P, Breyne P, Storme V, Boerjan W, van Slycken J (2006). Paternity analysis of Populus nigra L. offspring in a Belgian plantation of native and exotic poplars. Annals of Forest Science, 63, 783-790. DOI URL
[25]	Whitehead DR (1983). Wind pollination: some ecological and evolutionary perspectives. In: Real L ed. Pollination Biology. Academic Press, Orlando. 97-108.
[26]	Zhang DM ( 张冬梅), Shen XH ( 沈熙环), He TH ( 何田华) (2001). A paternity analysis of seeds from different clones in a Pinus tabulaeformis Carr. seed orchard. Acta Phytoecologica Sinica (植物生态学报), 25, 166-175. (in Chinese with English abstract)
[27]	Zhang DM ( 张冬梅), Sun PG ( 孙佩光), Shen XH ( 沈熙环), Ru GX ( 茹广欣) (2009). Paternity analysis of open- and control-pollinated seeds collected from a seed orchard of Pinus tabulaeformis. Journal of Plant Ecology (Chinese Version) (植物生态学报), 33, 302-310. (in Chinese with English abstract)
[28]	Zhang DM ( 张冬梅), Yang Y ( 杨娅), Shen XH ( 沈熙环), Ru GX ( 茹广欣) (2007). Selection of primers and establish- ment of SSR PCR reaction system on Pinus tabulaeformis Carr. Journal of Beijing Forestry University (北京林业大学学报), 29(2), 13-17. (in Chinese with English abstract)
[29]	Zhang W ( 张薇), Gong J ( 龚佳), Ji KS ( 季孔庶) (2009). Analyses on the mating system in seedling seed orchard of Pinus massoniana. Scientia Silvae Sinicae (林业科学), 45(6), 22-26. (in Chinese with English abstract)
[30]	Zhang ZW, Sui JJ, Mei L, Su CQ (2008). Pollen dispersal and its spatial distribution in a seed orchard of Larix kaempferi(Lamb.) Carr. Silvae Genetica, 57, 256-261.
[31]	Zhou Y, Bui T, Auckland LD, Williams CG (2002). Undermethylated DNA as a source of microsatellites from a conifer genome. Genome, 45, 91-99. URL PMID

位点 Locus	来源 Source	引物序列(5′-3′) Primer sequences (5′-3′)	重复单位 Repeat units	参考文献 Reference
PTest1	火炬松 Pinus taeda	F)CGATGTCGATTAGGGATTGG R)CCTGTTCTTCGTCGGATGTT	(GGA)₆	Zhang et al., 2009
PtTX4001	火炬松 P. taeda	F)CTATTTGAGTTAAGAAGGGAGTC R)CTGTGGGTAGCATCATC	(GT)₅	Zhou et al., 2002
RPTest11	火炬松 P. taeda	F)AGGATGCCTATGATATGCGC R)AACCATAACAAAAGCGGTCG	(CAT)₇	Zhang et al., 2009
PtTX2123	火炬松 P. taeda	F)AAGAACCCACAAACACAAG R)GGGCAAGAATTCAATGATAA	(AGC)₈	Zhang et al., 2009
PtTX3116	火炬松 P. taeda	F)CCTCCCAAAGCCTAAAGAAT R)CATACAAGGCCTTATCTTACAGAA	(TTG)₇…(TTG)₅	Ai et al., 2006
RPS160	北美乔松 P. strobus	F)ACTAAGAACTCTCCCTCTCACC R)TCATTGTTCCCCAAATCAT	(ACAG)₃AGGC(ACAC)₃	Ai et al., 2006
PtTX4062	火炬松 P. taeda	F)TCTAGGCAATCTTTTTACCAAC R)ATCATAGCCTCATCCAATACA	(T)₈(GT)₁₃	Zhou et al., 2002
PR203	辐射松 P. radiata	F)TGGGACCCCATATTCTGATG R)CATTCCACTAGTTCTCTCGCAC	(GA)₁₄	Zhang et al., 2007
PR4.6	辐射松 P. radiata	F)GAAAAAAAGGCAAAAAAAAGGAG R)ACCCAAGGCTACATAACTCG	(CA)₂₁(TA)₆	Zhang et al., 2007
PtTX4084	火炬松 P. taeda	F)ACTGGCGAAGGCGAGCCGACAC R)ACCGCTGTGGGAACCCTCCTCGTC	(GCT)₈	Zhou et al., 2002
Cjgssr170	日本柳杉 Cryptomeria japonica	F)TGGCGATGTGGTTCTTATGG R)CAACCGCATCTTTCCCTAATTC	(CT)₃₃	Ai et al., 2006

位点 Locus	来源 Source	引物序列(5′-3′) Primer sequences (5′-3′)	重复单位 Repeat units	参考文献 Reference
PTest1	火炬松 Pinus taeda	F)CGATGTCGATTAGGGATTGG R)CCTGTTCTTCGTCGGATGTT	(GGA)₆	Zhang et al., 2009
PtTX4001	火炬松 P. taeda	F)CTATTTGAGTTAAGAAGGGAGTC R)CTGTGGGTAGCATCATC	(GT)₅	Zhou et al., 2002
RPTest11	火炬松 P. taeda	F)AGGATGCCTATGATATGCGC R)AACCATAACAAAAGCGGTCG	(CAT)₇	Zhang et al., 2009
PtTX2123	火炬松 P. taeda	F)AAGAACCCACAAACACAAG R)GGGCAAGAATTCAATGATAA	(AGC)₈	Zhang et al., 2009
PtTX3116	火炬松 P. taeda	F)CCTCCCAAAGCCTAAAGAAT R)CATACAAGGCCTTATCTTACAGAA	(TTG)₇…(TTG)₅	Ai et al., 2006
RPS160	北美乔松 P. strobus	F)ACTAAGAACTCTCCCTCTCACC R)TCATTGTTCCCCAAATCAT	(ACAG)₃AGGC(ACAC)₃	Ai et al., 2006
PtTX4062	火炬松 P. taeda	F)TCTAGGCAATCTTTTTACCAAC R)ATCATAGCCTCATCCAATACA	(T)₈(GT)₁₃	Zhou et al., 2002
PR203	辐射松 P. radiata	F)TGGGACCCCATATTCTGATG R)CATTCCACTAGTTCTCTCGCAC	(GA)₁₄	Zhang et al., 2007
PR4.6	辐射松 P. radiata	F)GAAAAAAAGGCAAAAAAAAGGAG R)ACCCAAGGCTACATAACTCG	(CA)₂₁(TA)₆	Zhang et al., 2007
PtTX4084	火炬松 P. taeda	F)ACTGGCGAAGGCGAGCCGACAC R)ACCGCTGTGGGAACCCTCCTCGTC	(GCT)₈	Zhou et al., 2002
Cjgssr170	日本柳杉 Cryptomeria japonica	F)TGGCGATGTGGTTCTTATGG R)CAACCGCATCTTTCCCTAATTC	(CT)₃₃	Ai et al., 2006

位点 Locus	等位基因数目 Number of alleles	观测杂合度 Observed heterozygosity	期望杂合度 Expected heterozygosity	多态性信息含量 Polymorphic information content
PTest1	4	0.462	0.500	0.377
PtTX4001	6	0.291	0.321	0.292
RPTest11	2	0.467	0.359	0.294
PtTX2123	4	0.554	0.519	0.430
PtTX3116	9	0.758	0.841	0.820
RPS160	4	0.318	0.281	0.253
PtTX4062	4	0.272	0.236	0.209
PR203	8	0.568	0.691	0.637
PR4.6	11	0.598	0.605	0.563
PtTX4084	4	0.133	0.125	0.118
Cjgssr170	5	0.288	0.280	0.261
平均值 Mean	5.55	0.428	0.433	0.387

位点 Locus	等位基因数目 Number of alleles	观测杂合度 Observed heterozygosity	期望杂合度 Expected heterozygosity	多态性信息含量 Polymorphic information content
PTest1	4	0.462	0.500	0.377
PtTX4001	6	0.291	0.321	0.292
RPTest11	2	0.467	0.359	0.294
PtTX2123	4	0.554	0.519	0.430
PtTX3116	9	0.758	0.841	0.820
RPS160	4	0.318	0.281	0.253
PtTX4062	4	0.272	0.236	0.209
PR203	8	0.568	0.691	0.637
PR4.6	11	0.598	0.605	0.563
PtTX4084	4	0.133	0.125	0.118
Cjgssr170	5	0.288	0.280	0.261
平均值 Mean	5.55	0.428	0.433	0.387

父本 Male parent	交配母本 Female parent mated	子代数Numbers of progeny	繁殖贡献率Reproductive success (%)	父本 Male parent	交配母本 Female parent mated	子代数Numbers of progeny	繁殖贡献率Reproductive success (%)
1	31, 39	3	1.29	32	9, 19, 31, 52	6	2.59
2	19, 52	3	1.29	37	16, 39, 51, 52	6	2.59
3	31	2	0.86	38	39, 51	3	1.29
6	39, 51	3	1.29	39	16, 39, 51, 52	5	2.16
7	39, 51, 53	3	1.29	43	39, 52	2	0.86
11	16, 19	2	0.86	44	16, 31	5	2.16
12	9, 31, 39, 51, 53	9	3.88	48	16, 51, 52	3	1.29
13	16	2	0.86	49	16, 19, 52	5	2.16
14	9, 19, 31, 51	7	3.02	50	53	1	0.43
15	19, 39, 51	7	3.02	51	19	1	0.43
16	16, 19, 51, 52, 53	7	3.02	52	51	1	0.43
17	31, 51, 53	4	1.72	53	19	1	0.43
20	19, 52	2	0.86	54	9, 39, 51, 53	7	3.02
21	31, 53	2	0.86	55	9, 31, 39, 52	5	2.16
23	16, 19, 39, 51, 52	9	3.88	57	9, 16, 53	3	1.29
25	16, 19, 31, 52, 53	6	2.59	60	16, 19, 39	5	2.16
26	16, 39, 53	4	1.72	61	16, 39, 51, 53	12	5.17
28	9, 52, 53	3	1.29	63	19, 53	5	2.16
29	19, 31	4	1.72	66	16, 31, 52, 53	5	2.16
30	9, 16, 31, 51, 52, 53	8	3.45	62	9, 19, 39, 51, 53	6	2.59
31	9, 16, 31, 51, 53	7	3.02	88	16, 19, 39, 53	4	1.72
19	9, 16, 31, 39, 51, 52, 53	11	4.74	45	9, 16, 19, 31, 39, 53	8	3.45
47	16, 19, 31, 52, 53	10	4.31	58	9, 16, 19, 39, 52, 53	15	6.47