甘蔗抗褐锈病新基因抗感病池构建及SSR多态性引物筛选

doi:10.11924/j.issn.1000-6850.casb2020-0125

中国农学通报 ›› 2021, Vol. 37 ›› Issue (6): 97-103.doi: 10.11924/j.issn.1000-6850.casb2020-0125

所属专题：生物技术

甘蔗抗褐锈病新基因抗感病池构建及SSR多态性引物筛选

王晓燕(), 单红丽, 张荣跃, 仓晓燕, 王长秘, 李文凤, 尹炯, 罗志明, 黄应昆()

云南省农业科学院甘蔗研究所/云南省甘蔗遗传改良重点实验室,云南开远 661699

收稿日期:2020-05-25 修回日期:2020-07-26 出版日期:2021-02-25 发布日期:2021-02-24
通讯作者: 黄应昆
作者简介:王晓燕,女,1981年出生,云南开远人,副研究员,硕士,主要从事甘蔗病害研究。通信地址：661699 云南省开远市灵泉东路363号云南省农业科学院甘蔗研究所,Tel：0873-7227004,E-mail：xiaoyanwang402@sina.com。
基金资助:
国家自然科学基金项目“滇蔗茅对甘蔗褐锈病的抗性遗传规律分析及抗性基因定位”(31660419);国家现代农业产业技术体系(糖料)建设专项资金“甘蔗真菌性病害防控岗位科学家”(CARS-170303);云岭产业技术领军人才培养项目“甘蔗有害生物防控”(2018LJRC56);云南省技术创新人才培养对象项目(202005AD160012);云南省现代农业产业技术体系建设专项资金“云南省甘蔗产业技术体系植物保护与病害研究岗位科学家”(YNGZTX-4-92)

New Brown Rust Resistance Gene of Sugarcane: Construction of Resistant and Susceptible Gene Pools and SSR Polymorphic Primers Screening

Wang Xiaoyan(), Shan Hongli, Zhang Rongyue, Cang Xiaoyan, Wang Changmi, Li Wenfeng, Yin Jiong, Luo Zhiming, Huang Yingkun()

Sugarcane Research Institute, Yunnan Academy of Agricultural Sciences/Yunnan Key Laboratory of Sugarcane Genetic Improvement, Kaiyuan Yunnan 661699

Received:2020-05-25 Revised:2020-07-26 Online:2021-02-25 Published:2021-02-24
Contact: Huang Yingkun

摘要/Abstract

摘要：

由黑顶柄锈菌(Puccinia melanocephala H. Sydow ＆ P. Sydow)引起的甘蔗褐锈病是危害中国甘蔗生产的主要病害之一。为了鉴定和发掘抗褐锈病新基因,防止褐锈病爆发流行和保证甘蔗安全生产,本研究以杂交组合‘粤糖03-393’× ‘ROC 24’抗感分离真实性F₁代群体为材料,构建抗感基因池,合成449对引物对抗感亲本及抗感基因池进行抗、感连锁SSR分子标记筛选。结果表明,25对引物在抗感亲本间有多态性,其中4对引物(SMC236CG、SCESSR0928、SCESSR0636、SCESSR2551)在抗感亲本及抗感池间有多态性,初步判定这4个SSR标记在染色体上的位点可能与抗褐锈病新基因存在连锁关系。研究结果为后续开展抗褐锈病新基因定位、为中国甘蔗褐锈病防控及抗病育种奠定了良好的基础。

关键词: 甘蔗, 褐锈病, 抗病基因, 抗感病池, SSR标记

Abstract:

Brown rust caused by Puccinia melanocephala H. Sydow & P. Sydow is one of the main diseases that endanger sugarcane production in China. In order to identify and explore the new brown rust resistance gene, prevent the outbreaks of the disease and ensure the safe production of sugarcane, the authentic F₁ hybrid population of cross combination ‘Yuetang 03-393’× ‘ROC 24’ was used to construct resistant/susceptible gene pools, and then 449 pairs of primers were synthesized to screen SSR (simple-sequence repeat) markers that linked to resistance and susceptibility in the parental lines and the resistant/susceptible genetic pools in this study. The results showed that 25 pairs of primers were polymorphic between the parents, and among them four pairs of primers (SMC236CG, SCESSR0928, SCESSR0636, and SCESSR2551) were polymorphic between the parental lines and the resistant/susceptible genetic pools. It preliminarily indicated that the loci of these four SSR markers on chromosome might have linkage relationship with a new brown rust resistance gene. The results of this study establish a good foundation for the mapping of new brown rust resistance genes in the future, and for controlling sugarcane brown rust and breeding disease resistance varieties in China.

Key words: sugarcane, brown rust, resistant gene, resistant and susceptible gene pools, SSR marker

中图分类号:

S432.44

王晓燕, 单红丽, 张荣跃, 仓晓燕, 王长秘, 李文凤, 尹炯, 罗志明, 黄应昆. 甘蔗抗褐锈病新基因抗感病池构建及SSR多态性引物筛选[J]. 中国农学通报, 2021, 37(6): 97-103.

Wang Xiaoyan, Shan Hongli, Zhang Rongyue, Cang Xiaoyan, Wang Changmi, Li Wenfeng, Yin Jiong, Luo Zhiming, Huang Yingkun. New Brown Rust Resistance Gene of Sugarcane: Construction of Resistant and Susceptible Gene Pools and SSR Polymorphic Primers Screening[J]. Chinese Agricultural Science Bulletin, 2021, 37(6): 97-103.

图/表 3

参考文献 35

[1]	Rott P C, Girard J, Comstook J C. Impact of pathogen genetics on breeding for resistance to sugarcane diseases[J]. International Society of Sugar Cane Technologists Proceedings, 2013,28:1-11.
[2]	Purdy L H, Liu L J, Dean J L. Sugarcane rust: a newly important disease[J]. Plant Disease, 1983,67(11):1292-1296. doi: 10.1094/PD-67-1292 URL
[3]	Huang Y K, Li W F. Color Illustration of Diagnosis and Control for Modern Sugarcane Diseases, Pests, and Weeds[M]. China Agriculture Press, Beijing, China, 2018.
[4]	李文凤, 蔡青, 黄应昆, 等. 甘蔗野生资源对蔗茅柄锈菌的抗性鉴定[J]. 植物保护, 2005,31(2):51-53.
[5]	阮兴业, 杨雰, 孙楚坚. 云南省发现甘蔗蔗茅柄锈菌[J]. 真菌学报, 1983,2(04):260-261.
[6]	全国甘蔗重要病害研究协作组. 中国大陆植蔗省(区)(部分)甘蔗病害种类调查初报[J]. 甘蔗糖业, 1991(1):1-8.
[7]	韦金菊, 邓展云, 黄伟华, 等. 广西北海甘蔗锈病生物学特性研究及防治对策[J]. 安徽农业科学, 2010,38(27):14997-14999.
[8]	黄应昆, 李文凤. 云南蔗区甘蔗锈病流行原因及防治对策[J]. 植保技术与推广, 1998,18(5):22-23.
[9]	Raid R N, Comstock J C. Common rust. In: A guide to sugarcane diseases[M]. Rott P, Bailey RA, Comstock J C, Croft BJ, and Saumtally A S (Eds). La Librairie du Cirad, Montpellier, CIRAD and ISSCT, France. 2000: 85-89.
[10]	Daugrois J H, Grivet L, Roques D, et al. A putative major gene for rust resis-tance linked with a RFLP marker in sugarcane cultivar ‘R570’[J]. Theoretical and Applied Genetics, 1996,92:1059-1064. doi: 10.1007/BF00224049 URL pmid: 24166636
[11]	Asnaghi C, Roques D, Ruffel S, et al. Targeted mapping of a sugarcane rust resistance gene (Bru1) using bulked segregant analysis and AFLP markers[J]. Theoretical and Applied Genetics, 2004,108:759-764. doi: 10.1007/s00122-003-1487-6 URL
[12]	Cunff L L, Garsmeur O, Raboin L M, et al. Diploid/ polyploid syntenic shuttle mapping and haplotype-specific chromosome walking toward a rust resistance gene (Bru1) in highly polyploid sugarcane (2n approximately 12× approximately 115)[J]. Genetics, 2008,180:649-660. doi: 10.1534/genetics.108.091355 URL pmid: 18757946
[13]	Glynn N C, Laborde C, Davidson R W, et al. Utilization of a major brown rust resistance gene in sugarcane breeding[J]. Mol Breed, 2013,31:323-331.
[14]	Parco A S, Avellaneda M C, Hale A H, et al. Frequency and distribution of the brown rust resistance gene Bru1 and implications for the Louisiana sugarcane breeding programme[J]. Plant Breed, 2014,133:654-659.
[15]	李文凤, 王晓燕, 黄应昆, 等. 31份甘蔗野生核心种质资源褐锈病鉴定及Bru1基因的分子检测[J]. 作物学报, 2015,41:806-812. doi: 10.3724/SP.J.1006.2015.00806 URL
[16]	Li W F, Wang X Y, Huang Y K, et al. Molecular detection of Bru1 gene and identification of brown rust resistance in Chinese sugarcane germplasm[J]. Sugar Tech, 2016,19:183-190. doi: 10.1007/s12355-016-0453-y URL
[17]	Racedo J, Perera M F, Bertani R, et al. Bru1 gene and potential alternative sources of resistance to sugarcane brown rust disease[J]. Euphytica, 2013,191:429-436. doi: 10.1007/s10681-013-0905-3 URL
[18]	Glynn N C, Comstock J C, McCorkle K. Resistance to sugarcane brown rust: Opportunities and challenges in using the Bru1 resistance gene[C]// 10th pathology workshop of the International Society of Sugarcane Technologists, Nanning, China. 2012: 7-11.
[19]	Molina B L, Rosales-Longo F, Queme J L. Comparative analysis between phenotype and Bru1 marker for incidence to brown rust in sugarcane[J]. Proc. Int. Soc. Sugar Cane Technol, 2013,28:1-4.
[20]	Wang X Y, Li W F, Huang Y K, et al. Developing genetically segregating populations for localization of novel sugarcane brown rust resistance genes[J]. Euphytica, 2019. doi: 10.1007/s10681-017-2012-3 URL pmid: 32009665
[21]	Cordeiro G M, Taylor G O, Henry R J. Characterisation of microsatellite markers from sugarcane (Saccharum sp.), a highly polyploid species[J]. Plant Science, 2000,155:161-168. doi: 10.1016/s0168-9452(00)00208-9 URL pmid: 10814819
[22]	Pinto L R, Oliveira K M, Ulian E C, et al. Survey in the sugarcane expressed sequence tag database (SUCEST) for simple sequence repeats[J]. Genome, 2004,47:795-804. doi: 10.1139/g04-055 URL pmid: 15499394
[23]	Pan Y B. Highly polymorphic microsatellite DNA markers for sugarcane germplasm evaluation and variety identity testing[J]. Sugar Tech, 2006,8(4):246-256. doi: 10.1007/BF02943564 URL
[24]	Santos F R C, Pinto L R, Carlini-Garcia L A, et al. Marker-trait association and epistasis for brown rust resistance in sugarcane[J]. Euphytica, 2015,203:533-547.
[25]	Wang J P, Roe B, Macmi S, et al. Microcollinearity between autopolyploid sugarcane and diploid sorghum genomes[J]. BMC Genomics, 2010,11:261. doi: 10.1186/1471-2164-11-261 URL pmid: 20416060
[26]	Yang X P, Islam M S, Sushma S, et al. Identifying Quantitative Trait Loci (QTLs) and Developing Diagnostic Markers Linked to Orange Rust Resistance in Sugarcane (Saccharum spp.)[J]. Frontiers in Plant Science, 2018,9:1-10. doi: 10.3389/fpls.2018.00001 URL pmid: 29410674
[27]	李文凤, 王晓燕, 黄应昆, 等. 101份中国甘蔗主要育种亲本褐锈病抗性鉴定及Bru1基因的分子检测[J]. 作物学报, 2016,42(9):1411-1416. doi: 10.3724/SP.J.1006.2016.01411 URL
[28]	李文凤, 王晓燕, 黄应昆, 等. 34份甘蔗栽培原种抗褐锈病性鉴定及基因的分子检测[J]. 分子植物育种, 2015,13(8):1814-1821.
[29]	李竹, 许莉萍, 苏亚春, 等. 基于田间表型和Bru1基因检测分析甘蔗褐锈病抗性遗传[J]. 作物学报, 2018,4(2):306-312.
[30]	Kuar S, Panesar P S, Bear M B, et al. Simple sequence repeat markers in genetic divergence and marker-assisted selection of rice cultivars: a review[J]. Critical Reviews in Food Science & Nutrition, 2015,55(1):41-49. doi: 10.1080/10408398.2011.646363 URL pmid: 24915404
[31]	Chambers G K, Curtis C, Millar C D, et al. DNA fingerprinting in zoology: past, present, future[J]. Investigative Genetics, 2014,5(1):3. doi: 10.1186/2041-2223-5-3 URL pmid: 24490906
[32]	Kim K S, Sappington T W. Microsatellite data analysis for population genetics[J]. Methods in Molecular Biology, 2013,1006:271-295. doi: 10.1007/978-1-62703-389-3_19 URL pmid: 23546798
[33]	Raboin L M, Oliveira K M, Lecunff L, et al. Genetic mapping in sugarcane, a high polyploid, using bi-parental progeny: Identification of a gene controlling stalk color and a new rust resistance gene[J]. Theoretical & Applied Genetics, 2006,112:1382-1391 doi: 10.1007/s00122-006-0240-3 URL pmid: 16552554
[34]	张海英, 王振国, 毛爱军, 等. 与黄瓜白粉病抗病基因紧密连锁的SSR分子标记[J]. 华北农学报, 2008,23(6):77-80. doi: 10.7668/hbnxb.2008.06.017 URL
[35]	王坤, 王晓鸣, 朱振东, 等. 以SSR标记对普通菜豆抗炭疽病基因定位[J]. 作物学报, 2009,35(3):432-437. doi: 10.3724/SP.J.1006.2009.00432 URL

编号	引物		序列
72	SMC25DUQ		F: GCTTCCTAATCCATTGTTATTCTT R: GCCACTCCATCTGCTAGTGTTC
112	SC110M4-11a		F: CCACTGAGTTCCAAATCATGTCC; R: CGAGGATGTCTCCGAGGATG
155	SMC119CG		F: TTCATCTCTAGCCTACCCCAA R: AGCAGCCATTTACCCAGGA
196	SMC1760LA		F: CGCCAGGGAGATCAGCCA; R: CGTCCATTTTCAGATTGGGCTC
214	SMC2042FL		F: GGGTTCGATTCCCAGTCTG R: TTCTCGGAACGCATCTACC
215	SMC204CG		F: GCATTGGCGTCATCACTG R: TCCGTAACTGACCATTAAGAGC
230	SMC236CG		F: ACCGACCTCAAAGAAGACGGAT R: CAGTATGAGTGAAGCACGGGAAC
235	SMC24DUQ		F: CGCAACGACATATACACTTCGG R: CGACATCACGGAGCAATCAGT
237	SMC255CG		F: GATCGAGTAGGCAACAAAGCT R: CCAAAGTCACAACAGCCTCA
244	SMC286CS		F: TCAAATGGGACCTTATTGGAG R: TCCCTCGATCTCCGTTGTT
310	SMCI751CL		F: GCCATGCCCATGCTAAAGAT R: ACGTTGGTCCCGGAACCG
363	SCESSR0004		F: GTTTGGTTCCCGGAGGAGAG R: ACAATGGACACCTGAACGGC
364	SCESSR0390		F: ACAATGGACACCTGAACGGC R: GAGGAGAGAACCACAGCCGA
365	SCESSR0811		F: ACAATGGACACCTGAACGGC R: TCCCGGAGGAGATAACCACA
366	SCESSR1311		F: ACAATGGACACCTGAACGGC R: GTTTGGTTCCCGGAGGAGAG
367	SCESSR0939		F: CATTGCCCTCTCAAGCAACC R: TTTGCATCAGTCAGCCTGGA
376	MOLSSR2883		F: TGAGGTGTTGCCGTTTAATAGAGG R: ACGCGACATTCTTGAAGCTCAG
390	MOLSSR1707		F: GTGATCCCTTCCCAACGCTAC R: CACATCTCTCCTCGTCCAAAGACT
411		SCESSR0387		F: GACCAACACGGTGAACACGA R: CAGGCACGTCACCTCCTTCT
428		SCESSR0153		F: CCCGTTCGTCTCCAGCAG R: GTCGACGTGTGGGACTTCCT
435		SCESSR0928		F: CCACCGCAGCCGATATGA R: TTGAGAGGCTGACCCTGGAA
438		SCESSR2032		F: GACCTCGAGTGGTGGTTCCAT R: GTGCACCCAGTAGCACCCTC
439		MOLSSR2178		F: CGCTTTCGATTCGACTTCAACTT R: GTAGGGAGGGAAGGACCTGATG
443		SCESSR0636		F: TCCAACTTCATCCAAGTTTCCAA R: GCACCCACGAAGAGGATGAC
448		SCESSR2551		F: TGGACTAGGCCACCAGGAAA R: GTTTCGATGTGCTGCCTCCT

编号	引物		序列
72	SMC25DUQ		F: GCTTCCTAATCCATTGTTATTCTT R: GCCACTCCATCTGCTAGTGTTC
112	SC110M4-11a		F: CCACTGAGTTCCAAATCATGTCC; R: CGAGGATGTCTCCGAGGATG
155	SMC119CG		F: TTCATCTCTAGCCTACCCCAA R: AGCAGCCATTTACCCAGGA
196	SMC1760LA		F: CGCCAGGGAGATCAGCCA; R: CGTCCATTTTCAGATTGGGCTC
214	SMC2042FL		F: GGGTTCGATTCCCAGTCTG R: TTCTCGGAACGCATCTACC
215	SMC204CG		F: GCATTGGCGTCATCACTG R: TCCGTAACTGACCATTAAGAGC
230	SMC236CG		F: ACCGACCTCAAAGAAGACGGAT R: CAGTATGAGTGAAGCACGGGAAC
235	SMC24DUQ		F: CGCAACGACATATACACTTCGG R: CGACATCACGGAGCAATCAGT
237	SMC255CG		F: GATCGAGTAGGCAACAAAGCT R: CCAAAGTCACAACAGCCTCA
244	SMC286CS		F: TCAAATGGGACCTTATTGGAG R: TCCCTCGATCTCCGTTGTT
310	SMCI751CL		F: GCCATGCCCATGCTAAAGAT R: ACGTTGGTCCCGGAACCG
363	SCESSR0004		F: GTTTGGTTCCCGGAGGAGAG R: ACAATGGACACCTGAACGGC
364	SCESSR0390		F: ACAATGGACACCTGAACGGC R: GAGGAGAGAACCACAGCCGA
365	SCESSR0811		F: ACAATGGACACCTGAACGGC R: TCCCGGAGGAGATAACCACA
366	SCESSR1311		F: ACAATGGACACCTGAACGGC R: GTTTGGTTCCCGGAGGAGAG
367	SCESSR0939		F: CATTGCCCTCTCAAGCAACC R: TTTGCATCAGTCAGCCTGGA
376	MOLSSR2883		F: TGAGGTGTTGCCGTTTAATAGAGG R: ACGCGACATTCTTGAAGCTCAG
390	MOLSSR1707		F: GTGATCCCTTCCCAACGCTAC R: CACATCTCTCCTCGTCCAAAGACT
411		SCESSR0387		F: GACCAACACGGTGAACACGA R: CAGGCACGTCACCTCCTTCT
428		SCESSR0153		F: CCCGTTCGTCTCCAGCAG R: GTCGACGTGTGGGACTTCCT
435		SCESSR0928		F: CCACCGCAGCCGATATGA R: TTGAGAGGCTGACCCTGGAA
438		SCESSR2032		F: GACCTCGAGTGGTGGTTCCAT R: GTGCACCCAGTAGCACCCTC
439		MOLSSR2178		F: CGCTTTCGATTCGACTTCAACTT R: GTAGGGAGGGAAGGACCTGATG
443		SCESSR0636		F: TCCAACTTCATCCAAGTTTCCAA R: GCACCCACGAAGAGGATGAC
448		SCESSR2551		F: TGGACTAGGCCACCAGGAAA R: GTTTCGATGTGCTGCCTCCT

甘蔗抗褐锈病新基因抗感病池构建及SSR多态性引物筛选

New Brown Rust Resistance Gene of Sugarcane: Construction of Resistant and Susceptible Gene Pools and SSR Polymorphic Primers Screening

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