12个竹种的45S rDNA和5S rDNA分布特性比较

doi:10.11707/j.1001-7488.20220710

摘要/Abstract

摘要：

目的: 分析12个竹种的45S rDNA和5S rDNA分布特征，并调查这些竹种的染色体数目和结构，为竹类植物系统分类和染色体结构等研究提供相应的细胞遗传学资料，为相关的染色体识别提供相应的染色体物理标记。方法: 以毛竹、花毛竹、龟甲竹、早竹、金镶玉竹、白哺鸡竹、紫竹、斑竹、黄秆乌哺鸡竹、人面竹、大明竹和茶秆竹为材料，以45S rDNA和5S rDNA为探针，采用双色荧光原位杂交技术分析45S rDNA和5S rDNA在这些竹种染色体上的分布。结果: 1) 12个竹种的染色体数目均为2 n=48，45S rDNA在12个竹种染色体上展现出一定的多态性分布。①毛竹、花毛竹、龟甲竹、早竹、黄秆乌哺鸡竹和斑竹均具有2个45S rDNA位点，且2个45S rDNA位点信号强度和分布位置表现一致。②茶秆竹、人面竹、白哺鸡竹和金镶玉竹也具有2个45S rDNA位点，其中1个位点较为稳定，位于染色体末端，另外1个位点断裂严重，经常脱离分布位置。③紫竹和大明竹具有2个45S rDNA位点，这对位点在信号强度上存在较大差异，特别是紫竹表现更明显。2) 5S rDNA在12个竹种染色体上展现出了一定的多态性分布。①毛竹、花毛竹和龟甲竹的5S rDNA分布模式相似，具有2对信号强度不同的5S rDNA位点，其中1对5S rDNA位点信号很强，位于染色体末端，另外1对5S rDNA位点信号微弱，位于染色体近中间区域。②早竹、白哺鸡竹和黄秆乌哺鸡竹的5S rDNA分布模式较为相似，具有2对较强的5S rDNA信号，且2对信号强度差异不大，以成对形式位于2对同源染色体近中间区域。③金镶玉竹、紫竹和大明竹的5S rDNA分布模式比较特殊，均有3个位点，明显不对称，其中紫竹的3个5S rDNA位点，2个位于染色体末端，1个位于染色体近中间区域，金镶玉竹的3个5S rDNA，2个强弱不同的信号位于染色体中间，还有1个位于染色体末端，而大明竹的3个5S rDNA均位于染色体近中间区域。④斑竹与人面竹的5S rDNA分布模式非常相似，具有2对5S rDNA位点，且信号较弱，均位于染色体近中间区域。⑤茶秆竹具有1对较弱的5S rDNA位点，位于染色体短臂近着丝粒区域。结论: 1) 根据45S rDNA和5S rDNA在金镶玉竹、紫竹、人面竹、白哺鸡竹、大明竹和茶秆竹染色体上的分布特征，推测这些竹种的染色体可能发生过断裂、易位、删除或插入等遗传重组。2) 毛竹、花毛竹和龟甲竹3个竹种的45S rDNA和5S rDNA分布模式非常相似，该分布模式与3个竹种系统分类地位具有一定的一致性。

关键词: 竹类植物, 染色体, 荧光原位杂交, 5S rDNA, 45S rDNA

Abstract:

Objective: Bamboo is a facultative reproductive plant, with a large number of varieties and variants. Are there differences in chromosome number and structure among these bamboos? To solve these problems, the distribution characteristics of 45S rDNA and 5S rDNA of 12 different bamboo species was studied, meanwhile its chromosome number and structure was revealed, in order to provide corresponding chromosome physical markers for related chromosome recognition. Method: In this study, Phyllostachys edulis, P. edulis 'Tao kiang', P. edulis 'Heterocycla', P. violascens, P. aureosulcata 'Spectabilis', P. dulcis, P. nigra, P. reticulata 'Lacrima-deae', P. vivax f. aureocaulis, P. aurea, Pleioblastus gramineus and Pseudoasa amabilis were selected as materials, and 45S rDNA and 5S rDNA were used as probe. The distribution of 45S rDNA and 5S rDNA on the chromosomes of the 12 different bamboo species was studied by the two-color fluorescence in situ hybridization. Result: 1) The chromosome number of 12 bamboo species was 2 n=48, and their 45S rDNA distribution on the chromosomes showed a certain diversity. ①There were two 45S rDNA sites on the chromosome of P. edulis, P. edulis 'Tao kiang', P. edulis 'Heterocycla', P. violascens, P. vivax f. aureocaulis and P. reticulata 'Lacrima-deae', respectively, these 45S rDNA sites were consistent in distribution position and signal strength. ②There were also two 45S rDNA sites on the chromosomes of P. amabilis, P. aurea, P. dulcis and P. aureosulcata 'Spectabilis', but one of them was stable, located at the end of the chromosome, and the other site was seriously broken and often separated from the distribution position. ③Although P. nigra and P. gramineus also had two 45S rDNA sites, these two 45S rDNA differed greatly in signal strength, especially P. nigra, showing more obvious. 2) The 5S rDNA also showed a certain polymorphism distribution on the chromosomes of 12 bamboo species. ① P. edulis, P. edulis 'Tao kiang' and P. edulis 'Heterocycla' had similar 5S rDNA distribution with two pairs of 5S rDNA sites, of which one pair had strong signals and was located at the end of the chromosome, and another pair with weak signals was located near the middle region of the chromosome. ②P. violascens, P. dulcis and P. vivax f. aureocaulis had similar 5S rDNA distribution pattern, two pairs of 5S rDNA with similar fluorescence intensity were located near the middle region of the homologous chromosomes in pair. ③The distribution pattern of 5S rDNA of P. aureosulcata 'Spectabilis', P. nigra and P. gramineus was special, with three asymmetry 5S rDNA sites. As for P. nigra, two 5S rDNA sites were located at the end of chromosome in pair and another one was located near the middle region of a chromosome region alone. As for P. aureosulcata 'Spectabilis', two different 5S rDNA sites were located near the middle region of the chromosome, and another one was located at the end of chromosome alone. However, three 5S rDNA sites of P gramineus were all located near the middle region of the chromosome. ④The 5S rDNA distribution of P. reticulata 'Lacrima-deae' and P. aurea was similar, with two pairs of weak 5S rDNA signal sites that were located near the middle region of the chromosome. ⑤P. amabilis had a pair of 5S rDNA sites with weak signal, which were located near the centromere region of short arm of the chromosome. Conclusion: 1) According to the distribution characteristics of 45S rDNA and 5S rDNA of the chromosomes of P. aureosulcata 'Spectabilis', P. nigra, P. aurea, P. dulcis, P. gramineus and P. amabilis, we speculate that these bamboo species may have undergone chromosome rearrangement such as fracture, translocation, deletion or insertion. 2) P. edulis, P. edulis 'Tao kiang' and P. edulis 'Heterocycla'have similar 45S rDNA and 5S rDNA distribution pattern, which is consistent with their phylogenesis classification result.

Key words: bamboo plants, chromosome, Fluorescent in situ hybridization, 5S rDNA, 45S rDNA

中图分类号:

S718.43

徐川梅,王子晗,谢峰,金旭胤,吴心妍,陈荣,黄坚钦. 12个竹种的45S rDNA和5S rDNA分布特性比较[J]. 林业科学, 2022, 58(7): 93-102.

Chuanmei Xu,Zihan Wang,Feng Xie,Xuyin Jin,Xinyan Wu,Rong Chen,Jianqin Huang. Comparison of 45S rDNA and 5S rDNA Distribution Characteristics of 12 Bamboo Species[J]. Scientia Silvae Sinicae, 2022, 58(7): 93-102.

图/表 5

图1

表1

12个竹种的45S rDNA和5S rDNA分布特征"

竹种名称 Species	45S rDNA分布特点 Distribution of 45S rDNA	5S rDNA分布特点 Distribution of 5S rDNA
毛竹 P. edulis	1对末端信号 A pair of terminal signals	2对信号，1对强信号位于染色体末端，1对微弱信号位于另1对染色体近中间区域 Two pairs of signals, the pair of strong signals are located at the end of the chromosome, another pair of weak signals are located near the middle region of the chromosome
花毛竹 P. edulis ‘Tao kiang’	1对末端信号 A pair of terminal signals	2对信号，1对强信号位于染色体末端，1对微弱信号位于另1对染色体近中间区域 Two pairs of signals, the pair of strong signals are located at the end of the chromosome, another pair of weak signals are located near the middle region of the chromosome
龟甲竹 P. edulis ‘Heterocycla’	1对近末端信号 A pair of subterminal signals	2对信号，1对强信号位于染色体末端，1对微弱信号位于另1对染色体近中间区域 Two pairs of signals, the pair of strong signals are located at the end of the chromosome, another pair of weak signals are located near the middle region of the chromosome.
早竹 P. violascens	1对末端信号 A pair of terminal signal	2对较强的信号，均位于染色体近中间位置 Two pairs of strong signals are located near the middle region of the chromosome
白哺鸡竹 P. dulcis	1对末端信号, 其中一个位点发生了一定断裂 A pair of terminal signals, one of them is broken	2对较强的信号，均位于染色体近中间位置 Two pairs of strong signals are located near the middle region of the chromosome.
黄秆乌哺鸡竹P. vivax f. aureocaulis	1对末端信号，存在一定的断裂 A pair of broken terminal signals	2对信号，均位于染色体近中间位置 Two pairs of signals are located near the middle region of the chromosome
斑竹 P. reticulata ‘Lacrima-deae’	1对末端信号，存在一定的断裂 A pair of broken terminal signals	2对信号，均位于染色体近中间位置 Two pairs of signals are located near the middle region of the chromosome
人面竹 P. aurea	1对末端信号，其中一个位点发生了一定断裂 A pair of terminal signals, one of them is broken	2对信号，1对信号强度相似，另1对信号表现为1强1弱 Two pairs of signals, one pair are similar, another pair are different in intensity
金镶玉竹 P. aureosulcata ‘Spectabilis’	1对末端信号，其中一个位点发生了一定断裂 A pair of terminal signals, one of them is broken	3个信号，2个位于染色体近中间位置，表现1强1弱，1个位于染色体末端，信号较强 Three different signals, two of them are different in intensity and located near the middle region of the chromosome, the last one is strong and located at the end of the chromosome
紫竹 P. nigra	1强1弱2个信号，强信号易断裂 Two different strength signals, the strong one is easy to break.	3个信号，1对信号很强，位于染色体末端，另1个信号较弱，位于染色体近中间位置 Three signals, a strong pair are located at the end of the chromosome, another weak signal is located near the middle region of the chromosome
茶秆竹 P. amabilis	1对近末端信号，其中一个位点易断裂成2段 A pair of terminal signals, one of them easily breaks into 2 segments	1对较弱的信号，位于染色体短臂近着丝粒部位 A pair of weak signals are located near the centromere of the short arm
大明竹 P. gramineus	2个不同的末端信号 Two different terminal signals	3个信号，2个位于染色体短臂中间，另1个位于染色体近中间区域 Three signals, two are located at the middle of the short arm, the third located near the middle region of the chromosome

表1

图2

图3

图4

参考文献 0

	陈瑞阳, 李秀兰, 宋文芹, 等. 中国主要经济植物基因组染色体图谱Ⅳ. 北京: 科学出版社, 2003: 1- 628.
	Chen R Y , Li X L , Song W Q , et al. Chromosome atlas of major economic plants genome in china (Version IV): chromosome atlas of various bamboo species. Beijing: Science Press, Science Press, 2003: 1- 628.
	贾芳信, 周明兵, 陈荣, 等. 4种竹子的核型及其基因组大小. 林业科学, 2016, 52 (9): 57- 66.
	Jia F X , Zhou M B , Chen R , et al. Karyotype and genome size in four bamboo species. Scientia Silvae Sinicae, 2016, 52 (9): 57- 66.
	李秀兰, 林汝顺, 冯学琳, 等. 中国部分丛生竹类染色体数目报道. 植物分类学报, 2001, 39 (5): 433- 422.
	Li X L , Lin R S , Feng R S , et al. Chromosome number of some caespitose bamboos native in or introduced to China. Acta Phytotaxonomica Sinica, 2000, 39 (5): 433- 422.
	李秀兰, 刘松, 宋文芹, 等. 40种散生竹的染色体数目. 植物分类学报, 1999, 37 (6): 541- 544.
	Li X L , Liu S , Song W Q , et al. Chromosome number of forty species of scattered bamboos. Acta Phytotaxonomica Sinica, 1999, 37 (6): 541- 544.
	马乃训, 赖广辉, 张培新, 等. 中国刚竹属. 杭州: 浙江科学技术出版社, 2014: 28- 154.
	Ma N X , Lai G H , Zhang P X , et al. The genus Phyllostachys in China. Hangzhou: Zhejiang Science and Technology Press, 2014: 28- 154.
	徐川梅, 卢江杰, 汤定钦. 45S rDNA在7种竹子植物染色体上的定位. 林业科学, 2009a, 45 (12): 42- 45.
	Xu C M , Lu J J , Tang D Q . Physical mapping of the 45S rDNA on metaphase chromosomes in seven bamboo species by in situ hybridization. Scientia Silvae Sinicae, 2009a, 45 (12): 42- 45.
	徐川梅, 郑华威, 王骢, 等. 龟甲竹染色体C-分带、荧光原位杂交及其核型分析. 林业科学研究, 2009b, 22 (5): 691- 695.
	Xu C M , Zhen H W , Wang C , et al. Giemsa C-banding, FISH and karyotype analysis of Phyllostachys pubescens var.heterocycla chromosome. Forest Research, 2009b, 22 (5): 691- 695.
	张颖, 赵欣, 张圣虎, 等. 竹类植物修复重金属污染土壤的研究进展. 生态与农村环境学报, 2021, 37 (1): 30- 38.
	Zhang Y , Zhao X , Zhang S H , et al. Research progress of bamboo species in phytoremediation of heavy metal contaminated soil. Journal of Ecology and Rural Environment, 2021, 37 (1): 30- 38.
	Badaeva E D , Dedkova O S , Gay G , et al. Chromosomal rearrangements in wheat: their types and distribution. Genome, 2007, 50 (10): 907- 926. doi: 10.1139/G07-072
	Garcia S , Kovarík A , Leitch A R , et al. Cytogenetic features of rRNA genes across land plants: analysis of the plant rDNA database. Plant Journal, 2017, 89 (5): 1020- 1030. doi: 10.1111/tpj.13442
	Guo Z H , Ma P F , Yang G Q , et al. Genome sequences provide insights into the reticulate origin and unique traits of woody bamboos. Molecular Plant, 2019, 12 (10): 1353- 1365. doi: 10.1016/j.molp.2019.05.009
	Hasterok R , Wolny E , Hosiawa M , et al. Comparative analysis of rDNA distribution in chromosomes of various species of Brassicaceae. Annals of Botany, 2006, 97 (2): 205- 216. doi: 10.1093/aob/mcj031
	Heslop-Harrison J S . Comparative genome organization in plants: from sequence and markers to chromatin and chromosomes. Plant Cell, 2000, 12 (5): 617- 636. doi: 10.1105/tpc.12.5.617
	Huang J , Ma L , Yang F , et al. 45S rDNA regions are chromosome fragile sites expressed as gaps in vitro on metaphase chromosomes of root-tip meristematic cells in Lolium spp. PLoS One, 2008, 3 (5): e2167.
	Jiang J , Gill B S , Wang G L , et al. Metaphase and interphase fluorescence in situ hybridization mapping of the rice genome with bacterial artificial chromosomes. Proceedings of the National Academy of Sciences, 1995, 92 (10): 4487- 4491.
	Katsiotis A , Loukas M , Heslop-Harrison J S . Repetitive DNA, genome and species relationships in Avena and Arrhenatherum (Poaceae). Annals of Botany, 2000, 86 (6): 1135- 1142.
	Luo X M , Liu J C . Fluorescence in situ hybridization (FISH) analysis of the locations of the oligonucleotides 5S rDNA, (AGGGTTT)3, and (TTG) 6 in three genera of Oleaceae and their phylogenetic framework. Genes, 2019, 10 (5): 375.
	Mondin M , Aguiar-Perecin , M L R . Heterochromatin patterns and ribosomal DNA loci distribution in diploid and polyploid Crotalaria species (Leguminosae, Papilionoideae) and inferences on karyotype evolution. Genome, 2011, 54 (9): 718- 726.
	Monkheang P , Chaveerach A , Sudmoon R , et al. Karyotypic features including organizations of the 5S, 45S rDNA loci and telomeres of Scadoxus multiflorus (Amaryllidaceae). Comparative Cytogenetics, 2016, 10 (4): 637- 646.
	Osuji J O , Crouch J , Harrison G , et al. Molecular cytogenetics of Musa species, cultivars and hybrids: location of 18S-5.8S-25S and 5S rDNA and telomere-like sequences. Annals of Botany, 1998, 82 (2): 243- 248.
	Pedrosa A , Sandal N , Stougaard J , et al. Chromosomal map of the model legume Lotus japonicas. Genetics, 2002, 161 (4): 1661- 1672.
	Pedrosa-Harand A , de Almeida C C S , Mosiolek M , et al. Extensive ribosomal DNA amplification during Andean common bean (Phaseolus vulgaris L.) evolution. Theoretical and Applied Genetics, 2006, 112 (5): 924- 33.
	Peng Z , Lu Y , Li L , et al. The draft genome of the fast-growing non-timber forest species moso bamboo (Phyllostachys heterocycla). Nature Genetics, 2013, 45 (4): 456- 461.
	Raskina O , Barber J C , Nevo E , et al. Repetitive DNA and chromosomal rearrangements: speciation related events in plant genomes. Cytogenetic and Genome Research, 2008, 120 (3/4): 351- 357.
	Tsang E , Carr A M . Replication fork arrest, recombination and the maintenance of ribosomal DNA stability. DNA Repair, 2008, 7 (10): 1613- 1623.
	Vaio M , Speranza P , Valls J F , et al. Localization of the 5S and 45S rDNA sites and cpDNA sequence analysis in species of the Quadrifaria group of Paspalum (Poaceae, Paniceae). Annals of Botany, 2005, 96 (2): 191- 200.
	Vasconcelos E V , Vasconcelos S , Ribeiro T , et al. Karyotype heterogeneity in Philodendron s.l. (Araceae) revealed by chromosome mapping of rDNA loci. PLoS One, 2018, 13 (11): 1- 15.
	Zhang L N , Ma P F , Zhan Y X , et al. Using nuclear loci and allelic variation to disentangle the phylogeny of Phyllostachys (Poaceae, Bambusoideae). Molecular Phylogenetics and Evolution, 2019, 137, 222- 235.
	Zhang Z T , Yang S Q , Li Z A , et al. Comparative chromosomal localization of 45S and 5S rDNAs and implications for genome evolution in Cucumis. Genome, 2016, 59 (7): 449- 457.
	Zhao H , Gao Z , Wang L , et al. Chromosome-level reference genome and alternative splicing atlas of moso bamboo (Phyllostachys edulis). Gigascience, 2018, 7 (10): 1- 12.
	Zhou M B , Xu C M , Shen L F , et al. Evolution of genome sizes in Chinese Bambusoideae (Poaceae) in relation to karyotype. Trees, 2017, 31, 41- 48.

[1]	李艳民,袁德义,叶天文,陈雅,韩春霞,肖诗鑫. 油茶种间杂交F₁代18个优良单株核型分析[J]. 林业科学, 2022, 58(4): 165-174.
[2]	王律几,张晓晓,王君. 基于45S rDNA-FISH的异源三倍体'银中杨’小孢子母细胞减数分裂染色体行为追踪[J]. 林业科学, 2022, 58(3): 69-78.
[3]	叶天文,袁德义,李艳民,肖诗鑫,龚守富,张健,李素芳,罗健. 海南油茶的倍性鉴定[J]. 林业科学, 2021, 57(7): 61-69.
[4]	郭瑞红,邱帅,刘光欣,高凯,魏建芬,席梦利. 绣球染色体制片技术优化及rDNA物理定位[J]. 林业科学, 2021, 57(11): 59-67.
[5]	许自龙, 陈益存, 高暝, 吴立文, 赵耘霄, 汪阳东. 被子植物性别分化的研究进展[J]. 林业科学, 2019, 55(8): 157-169.
[6]	王吉锐, 郑晨义, 杜予州, 王浩, 周国鑫, 徐志宏. 危害竹子的3种粉虱记述(半翅目:粉虱科)[J]. 林业科学, 2018, 54(7): 173-180.
[7]	田梦迪, 李燕杰, 张平冬, 王健, 郝静颐. 高温诱导银灰杨花粉染色体加倍创制杂种三倍体[J]. 林业科学, 2018, 54(3): 39-47.
[8]	徐川梅, 黄坚钦, 王正加, 夏国华, 张启香, 黄有军, 张守攻. 山核桃花粉母细胞减数分裂及核型[J]. 林业科学, 2017, 53(11): 77-84.
[9]	贾芳信, 周明兵, 陈荣, 杨海芸, 高培军, 徐川梅. 4种竹子的核型及其基因组大小[J]. 林业科学, 2016, 52(9): 57-66.
[10]	于存, 池玉杰. 一色齿毛菌锰过氧化物酶（MnP）活性检测与MnP1基因的克隆[J]. 林业科学, 2014, 50(3): 109-116.
[11]	江泽慧. 竹类植物基因组学研究进展[J]. 林业科学, 2012, 48(1): 159-166.
[12]	徐川梅卢江杰汤定钦. 45S rDNA在7种竹子植物染色体上的定位*[J]. 林业科学, 2009, 12(12): 42-45.
[13]	王小蓉汤浩茹付华清钟必凤夏武峰唐海东邓群仙. 15 个引进树莓品种的核型[J]. 林业科学, 2008, 44(12): 147-150.
[14]	李明银. 马蹄纹天竺葵异倍型嵌合体不同层体细胞染色体数目的直接测定(英文)[J]. 林业科学, 2007, 43(7): 125-127.
[15]	马乃训陈光才袁金玲. 国产竹类植物生物多样性及保护策略[J]. 林业科学, 2007, 43(4): 102-106.