利用Target SSR-seq技术鉴定温州蜜柑芽变材料

doi:10.3864/j.issn.0578-1752.2021.23.013

中国农业科学 ›› 2021, Vol. 54 ›› Issue (23): 5083-5096.doi: 10.3864/j.issn.0578-1752.2021.23.013

利用Target SSR-seq技术鉴定温州蜜柑芽变材料

胡冬梅¹(),江东²(),李永平³,彭磊⁴(),李冬云⁵,朱延松²,杨云光¹

¹玉溪市农业技术推广站,云南玉溪 653100
²西南大学柑桔研究所,重庆 400712
³云南省绿色食品发展中心,昆明 650032
⁴云南农业大学园林园艺学院,昆明 650201
⁵华宁县柑桔产业发展办公室,云南华宁 652800

收稿日期:2021-02-22 接受日期:2021-05-20 出版日期:2021-12-01 发布日期:2021-12-06
通讯作者: 江东,彭磊
作者简介:胡冬梅,Tel：15008875127;E-mail： 909625192@qq.com。
基金资助:
国家重点研发计划(2018YFD1000101);云南省重大科技专项—绿色食品国际合作研究中心项目(2019ZG00907)

Identification of Bud Sport Mutation of Satsuma Mandarin by Target SSR-seq Technology

HU DongMei¹(),JIANG Dong²(),LI YongPing³,PENG Lei⁴(),LI DongYun⁵,ZHU YanSong²,YANG YunGuang¹

¹Agricultural Technology Extension Station of Yuxi County, Yuxi 653100, Yunnan
²Citrus Research Institute of Southwest University, Chongqing 400712
³Yunnan Green Food Development Center, Kunming 650032
⁴College of Landscape and Horticulture, Yunnan Agricultural University, Kunming 650201
⁵Citrus Industry Development Office of Huaning County, Huaning 652800, Yunnan

Received:2021-02-22 Accepted:2021-05-20 Online:2021-12-01 Published:2021-12-06
Contact: Dong JIANG,Lei PENG

摘要/Abstract

摘要：

【目的】柑橘芽极易发生变异,用形态学的方法鉴定这些芽变材料,易受环境、栽培条件等因素影响,而利用深度测序技术开展柑橘芽变材料鉴定,能够获得变异位点的基因型;同时,建立的技术体系也有利于柑橘资源的精准鉴定、种质资源的遗传多样性研究和植物性品种权保护。【方法】本研究通过靶位点SSR测序技术对柑橘芽变材料开展鉴定。首先利用柑橘全基因组序列扫描发掘SSR,采用多态性较高的SSR位点用于柑橘芽变材料的区分,进一步利用多路复用PCR扩增构建SSR靶位点文库,并通过MiSeq深度测序方法对2份温州蜜柑芽变材料的SSR靶位点进行测序验证。再利用开发的SSR标记对22份优质柑橘种质资源进行遗传多样性分析。【结果】利用GMATA软件在克里曼丁红橘（Citrus clementina Hort. ex. Tanaka）参考基因组和温州蜜柑（Citrus unshiu Macf.）基因组中分别获得69 101个和80 193个SSR,其中以AT/TA为主的2基序SSR最多,3基序中以AAT最多,通过序列比对发掘出变异热点区域的高多态性SSR用于温州蜜柑芽变材料的检测,4对SSR引物能对22份柑橘材料进行准确区分。77对SSR靶位点的深度测序一次性对多个SSR位点进行基因分型,能准确鉴定出2份温州蜜柑芽变材料的SSR基因型,以及SSR在基因组上的准确位置。结合SSR以及SNP基因型,能够对2份温州蜜柑芽变材料进行有效区分。【结论】本研究开发了用于柑橘芽变材料区分的高效SSR位点发掘方法,结合靶位点多路扩增和Target SSR-seq的深度测序实现了柑橘芽变材料的高效区分。

关键词: 柑橘, 芽变, Target SSR-seq, 深度测序

Abstract:

【Objective】 Most citrus cultivars especially some seedless varieties were originated from bud sports mutation. The plant morphological traits of these bud sports were susceptible to environment and cultivation conditions, so discriminating the mutant lines from its parents have been still hard tasks although many molecular marker approaches were developed. As the use of deep sequencing technology could simultaneously genotype multiple loci of the mutation lines, it was already widely used to discriminate citrus bud sports in high efficiency. In addition, the technical approach could also benefit to accurate the identification of citrus genetic accessions, the genetic diversity research and the variety rights protection. 【Method】 In this study, the Target-SSR sequencing approach was adopted to discriminate citrus bud sport. The SSR loci were firstly discovered by scanning of the clementine mandarin (Citrus reticulata Blanco) reference genome and satsuma mandarin (Citrus unshiu Macf.) genome sequences, and those high polymorphic SSR loci were used to design primers to distinguish 22 citrus accessions which including some bud sports materials. Finally, multiplexed PCR production obtained through 18 runs PCR amplification with 77 pair primers on two bud sports materials were used to construct target SSR sequencing library, and sequenced with MiSeq apparatus to genotype SSR and SNP variation. In addition, the genetic diversity of 22 citrus accessions were also analyzed by using the developed SSR markers in this study. 【Result】 69 101 SSRs and 80 193 SSR were obtained respectively from clementine reference genome and satsuma genome by using GMATA software, among which AT/TA motif SSR were the most, and 3 motifs AAT rank the second. The high polymorphic SSR loci and its flanking sequences were extracted to design primers for bud sports discrimination and library construction. Four pairs of SSR primers could accurately distinguish 22 citrus germplasms. Target SSR-sequencing with MiSeq apparatus not only simultaneously genotype multiple SSR loci at one time but also accurately identified the variant loci of SSRs. Combining with the identified SSR and SNP genotypes, it could be effectively distinguished two satsuma bud sports. 【Conclusion】In this research, an efficient SSR loci discovering method was developed, combining with target SSR multiplex amplification and deep sequencing on MiSeq platform, which could effectively discriminate citrus bud sports. This study not only provided an important means to identify citrus buds sports, but also facilitated the protection of citrus varieties and the management of citrus germplasms genetic diversity in the future.

Key words: citrus, bud sports, Target SSR-seq, deep sequencing

胡冬梅,江东,李永平,彭磊,李冬云,朱延松,杨云光. 利用Target SSR-seq技术鉴定温州蜜柑芽变材料[J]. 中国农业科学, 2021, 54(23): 5083-5096.

HU DongMei,JIANG Dong,LI YongPing,PENG Lei,LI DongYun,ZHU YanSong,YANG YunGuang. Identification of Bud Sport Mutation of Satsuma Mandarin by Target SSR-seq Technology[J]. Scientia Agricultura Sinica, 2021, 54(23): 5083-5096.

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参考文献 36

[1]	董美超, 李进学, 周东果, 岳建强, 高俊燕. 柑橘品种选育研究进展. 中国果树, 2013(6):73-78.
	DONG M C, LI J X, ZHOU D G, YUE J Q, GAO J Y. Advances in citrus breeding. China fruit, 2013(6):73-78. (in Chinese)
[2]	邓秀新, 王力荣, 李绍华, 张绍铃, 张志宏, 丛佩华, 易干军, 陈学森, 陈厚彬, 钟彩虹. 果树育种40年回顾与展望. 果树学报, 2019, 36(4):514-520.
	DENG X X, WANG L R, LI S H, ZHANG S L, ZHANG Z H, CONG P H, YI G J, CHEN X S, CHEN H B, ZHONG C H. Retrospection and prospect of fruit breeding for last four decades in China. Journal of Fruit Science, 2019, 36(4):514-520. (in Chinese)
[3]	张敏, 邓秀新. 柑橘芽变选种以及芽变性状形成机理研究进展. 果树学报, 2006, 23(6):871-876.
	ZHANG M, DENG X X. Advances in research of Citrus cultivars selected by bud mutation and the mechanism of formation of mutated characteristics. Journal of Fruit Science, 2006, 23(6):871-876. (in Chinese)
[4]	王三红, 陈力耕, 章镇, 房经贵. RAPD在柑橘品系鉴别上的应用. 果树科学, 2000, 17(1):70-72.
	WANG S H, CHEN L G, ZHANG Z, FANG J G. Application of RAPD in identification of Citrus strains. Journal of Fruit Science, 2000, 17(1):70-72. (in Chinese)
[5]	范眸天, 高俊, 吴兴恩, 李文祥, 龙雯虹, 许明辉. 十五种柑桔种质资源的RAPD分析. 中国南方果树, 2002, 31(6):3-6.
	FAN M T, GAO J, WU X E, LI W X, LONG W H, XU M H. The RAPD analysis of fifteen germplasm resources of Citrus. South China Fruits, 2002, 31(6):3-6. (in Chinese)
[6]	罗静, 周心智, 张云贵. RAPD标记对25份柑桔资源及其芽变系的鉴定和多样性分析. 中国农学通报, 2008, 24(8):99-104.
	LUO J, ZHOU X Z, ZHANG Y G. Identification and genetic diversity analysis of 25 Citrus resources and its sports strains by RAPD technique. Chinese Agricultural Science Bulletin, 2008, 24(8):99-104. (in Chinese)
[7]	吴兴恩, 范眸天, 龚洵, 杨杨. 22份柑桔资源的ISSR分析. 云南农业大学学报, 2006, 21(1):36-41, 51.
	WU X E, FAN M T, GONG X, YANG Y. The ISSR analysis of 22 Citrus resources. Journal of Yunnan Agricultural University, 2006, 21(1):36-41, 51. (in Chinese)
[8]	刘通, 邓崇岭, 程玉芳, 李秋景, 陈传武, 刘冰浩, 伊华林. 利用SSR 和 SRAP 技术分析广西柑橘种质, 遗传多样性. 2016, 35(2):23-29.
	LIU T, DENG C L, CHENG Y F, LI Q J, CHEN C W, LIU B H, YI H L. Genetic diversity analysis of Citrus germplasm in Guangxi using SSR and SRAP techniques. Journal of Huazhong Agricultural University, 2016, 35(2):23-29. (in Chinese)
[9]	BI Q X, ZHAO Y, CUI Y F, WANG L B. Genome survey sequencing and genetic background characterization of yellow horn based on next-generation sequencing. Molecular Biology Reports, 2019, 46(4):4303-4312. doi: 10.1007/s11033-019-04884-7
[10]	韩国辉. 基于EST-SSR、Genomic-SSR 和SCoT 标记的柑橘连锁图谱构建及杂种和多倍体遗传分析[D]. 重庆: 西南大学, 2012.
	HAN G H. Construction of linkage map and genetic analysis of hybrid and polyploid in citrus based on EST-SSR, Genomic-SSR and Scot markers[D]. Chongqing: Southwest University, 2012. (in Chinese)
[11]	李益, 马先锋, 唐浩, 李娜, 江东, 龙桂友, 李大志, 牛英, 韩瑞玺, 邓子牛. 柑橘品种鉴定的SSR标记开发和指纹图谱库构建. 中国农业科学, 2018, 51(15):2969-2979.
	LI Y, MA X F, TANG H, LI N, JIANG D, LONG G Y, LI D Z, NIU Y, HAN R X, DENG Z N. SSR markers screening for identification of Citrus cultivar and construction of DNA fingerprinting library. Scientia Agricultura Sinica, 2018, 51(15):2969-2979. (in Chinese)
[12]	曹庆芹, 孟海军, 文晓鹏, 伊华林, 邓秀新. 利用SSR标记分析柑橘雄性不育及低育资源的遗传多样性. 农业生物技术学报, 2006, 14(6):937-941.
	CAO Q Q, MENG H J, WEN X P, YI H L, DENG X X. Genetic diversity of male sterile and low fertile germplasm of Citrus reealed using SSR markers. Journal of Agricultural Biotechnology, 2006, 14(6):937-941. (in Chinese)
[13]	YAMAMOTO T, MOCHIDA K, IMAI T, HAJI T, YAEGAKI H, YAMAGUCHI M, MATSUTA N, OGIWARA I, HAYASHI T. Parentage analysis in Japanese peaches using SSR markers. Breeding Science, 2003, 53(1):35-40. doi: 10.1270/jsbbs.53.35
[14]	徐云碧, 杨泉女, 郑洪建, 许彦芬, 桑志勤, 郭子锋, 彭海, 张丛, 蓝昊发, 王蕴波, 吴坤生, 陶家军, 张嘉楠. 靶向测序基因型检测(GBTS)技术及其应用. 中国农业科学, 2020, 53(15):2983-3004.
	XU Y B, YANG Q N, ZHENG H J, XU Y F, SANG Z Q, GUO Z F, PENG H, ZHANG C, LAN H F, WANG Y B, WU K S, TAO J J, ZHANG J N. Genotyping by target sequencing (GBTS) and its applications. Scientia Agricultura Sinica, 2020, 53(15):2983-3004. (in Chinese)
[15]	黄卫, 罗玉萍. DNA分子标记在柑桔中的应用. 生物技术, 2002, 12(1):34-36.
	HUANG W, LUO Y P. Application of DNA molecular marker in Citrus. Biotechnology, 2002, 12(1):34-36. (in Chinese)
[16]	MICHAELSON J J, SHI Y J, GUJRAL M, ZHENG H C, MALHOTRA D, JIN X, JIAN M H, LIU G M, GREER D, BHANDARI A, WU W T, COROMINAS R, PEOPLES A, KOREN A, GORE A, KANG S L, LIN G N, ESTABILLO J, LAKOUCHEVA L M, LI Y R, WANG J, SEBAT J. Whole-genome sequencing in autism identifies hot spots for de novo germline mutation. Cell, 2012, 151(7):1431-1442. doi: 10.1016/j.cell.2012.11.019
[17]	SOHN H B, KIN S J, HWANG T Y, PARK H M, LEE Y Y, MARKKANDAN K, LEE D, LEE S, HONG S Y, SONG Y H, KOO B C, KIM Y H. Barcode system for genetic identification of soybean (Glycine max(L.) Merrill) cultivars using Indel markers specific to dense variation blocks. Frontiers in Plant Science, 2017, 8:520.
[18]	LI L, FANG Z W, ZHOU J F, CHEN H, HU Z F, GAO L F, CHEN L H, REN S, MA H Y, LU L, ZHANG W X, PENG H. An accurate and efficient method for large-scale SSR genotyping and applications. Nucleic Acids Research, 2017, 45(10):e88. doi: 10.1093/nar/gkx093
[19]	刘欢, 黄丽娜, 张奋强, 姜梦嫣, 周义杰, 陈思源, 王有年, 杨明峰, 马兰青. RNA-Seq高通量测序技术在果树功能基因组学研究的应用进展. 生物技术进展, 2017, 7(2):144-148.
	LIU H, HUANG L N, ZHANG F Q, JIANG M Y, ZHOU Y J, CHEN S Y, WANG Y N, YANG M F, MA L Q. Progress on application of RNA-seq high throughput sequencing technology in functional genomics of fruit trees. Current Biotechnology, 2017, 7(2):144-148. (in Chinese)
[20]	张先文, 贺治洲, 江南, 邓华凤, 李继明. 高通量基因型分型技术及其在水稻中的应用. 生物技术通报, 2017, 33(12):67-73.
	ZHANG X W, HE Z Z, JIANG N, DENG H F, LI J M. High-throughput genotyping techniques and their applications in rice. Biotechnology Bulletin, 2017, 33(12):67-73. (in Chinese)
[21]	YANG J J, ZHANG J, HAN R X, ZHANG F, MAO A J, LUO J, DONG B B, LIU H, TANG H, ZHANG J N, WEN C L. Target SSR-seq: A novel SSR genotyping technology associate with perfect SSRs in genetic analysis of cucumber varieties. Frontiers in Plant Science, 2019, 10:531. doi: 10.3389/fpls.2019.00531
[22]	LI H, MA Y S, PENG F Y, ZHANG H Y, LIU J C, JIANG M. Large-scale advances in SSR markers with high-throughput sequencing in Euphorbia fischeriana Steud. Electronic Journal of Biotechnology, 2021, 49(1):50-55. doi: 10.1016/j.ejbt.2020.11.004
[23]	FORDYCE S L, ávila-Arcos M C, ROCKENBAUER E, BORSTING C, FRANK-HANSEN R, PETERSEN F T, WILLERSLEV E, HANSEN A J, MORLING N, GILBERT M T P. High-throughput sequencing of core STR loci for forensic genetic investigations using the Roche Genome Sequencer FLX platform. BioTechniques, 2011, 51:127-133. doi: 10.2144/000113721
[24]	谢让金, 邓烈. 一种适合AFLP分析的柑橘DNA提取方法. 生物技术, 2007, 17(6):27-28.
	XIE R J, DENG L. A Citrus DNA extraction method for AFLP analysis. Biotechnology, 2007, 17(6):27-28. (in Chinese)
[25]	XANG X W, WANG L. GMATA: An integrated software package for genome-scale SSR mining, marker development and viewing. Frontiers in Plant Science, 2016, 7:1350.
[26]	UNTERGASSER A, CUTCUTACHE I, KORESSAAR T, YE J A, FAIRCLOTH B C, REMM M, ROZEN S G. Primer3-new capabilities and interfaces. Nucleic Acids Research, 2012, 40(15):e115. doi: 10.1093/nar/gks596
[27]	KALINOWSKI S T, TAPER M L, MARSHALL T C. Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Molecular Ecology, 2007, 16(5):1099-1106. doi: 10.1111/j.1365-294X.2007.03089.x
[28]	PEAKALL R, SMOUSE P E. GENALEX 6: Genetic analysis in Excel. Population genetic software for teaching and research. Molecular Ecology Notes, 2006, 6(1):288-295. doi: 10.1111/men.2006.6.issue-1
[29]	THORVALDSDÓTTIR H, ROBINSON J T, MESIROV J P. Integrative Genomics Viewer (IGV): High-performance genomics data visualization and exploration. Briefings in Bioinformatics, 2012, 14(2):178-192. doi: 10.1093/bib/bbs017
[30]	孙淑霞, 陈栋, 李靖, 涂美艳, 谢红江, 江国良. 北京28号桃芽变株系的ISSR和SSR鉴定. 果树学报, 2012, 29(1):24-28.
	SUN S X, CHEN D, LI J, TU M Y, XIE H J, JIANC G L. Molecular identification of peach bud sports from Beijing 28 with ISSR and SSR. Journal of Fruit Science, 2012, 29(1):24-28. (in Chinese)
[31]	齐丹. 梨种质变异的SRAP和EST-SSR分析[D]. 北京: 中国农业科学院, 2013.
	QI D. Analysis of variation of pear germplasm accessions by SRAP and EST-SSR markers[D]. Beijing: Chinese Academy of Agricultural Sciences, 2013. (in Chinese)
[32]	LI L, FANG Z W, ZHOU J F, CHEN H, HU Z F, GAO L F, CHEN L H, REN S, MA H Y, LU L, ZHANG W X, HAI P. An accurate and efficient method for large-scale SSR genotyping and applications. Nucleic Acids Research, 2017, 45(10)V: e88.
[33]	MA J J, ZHAO Y H, CHEN H, FU C, ZHU L, ZHOU X M, XIA H, HOU L, LI G G, ZHUANG W J, WANG X J, ZHAO C Z. Genome-wide development of polymorphic microsatellite markers and their application in peanut breeding program. Electronic Journal of Biotechnology, 2020, 44(3):25-32. doi: 10.1016/j.ejbt.2020.01.004
[34]	WANG Y, YANG C, JIN Q J, ZHOU D J, WANG S S, YU Y J, YANG L. Genome-wide distribution comparative and composition analysis of the SSRs in Poaceae. BMC Genet, 2015, 16(2):18. doi: 10.1186/s12863-015-0178-z
[35]	QI W H, LU T, ZHENG C L, JIANG X M, JIE H, ZHANG X Y, YUE B S, ZHAO G J. Distribution patterns of microsatellites and development of its marker in different genomic regions of forest musk deer genome based on high throughput sequencing. Aging, 2020, 12(5):4445-4462. doi: 10.18632/aging.v12i5
[36]	PAVAN KUMAR P, JANAKIRAM T, BHAT K V. Microsatellite based DNA fingerprinting and assessment of genetic diversity in Bougainvillea cultivars. Gene, 2020, 753:144794. doi: 10.1016/j.gene.2020.144794

序号 Code	种质号 Germplasm number	采集地 Collection site	对应母本 Corresponding maternal parent	序号 Code	种质号 Germplasm number	采集地 Collection site	对应母本 Corresponding maternal parent
1	芽变1号 Bud sports No1.	华宁县 Huaning county	4	12	芽变5号 Bud sports No5.	华宁县 Huaning county	11
2	芽变2号 Bud sport No2.	华宁县 Huaning county	4	13	大分1号 Oita No1.	华宁县 Huaning county
3	芽变3号 Bud sport No3.	华宁县 Huaning county	9	14	HX10	华宁县 Huaning county
4	冰糖橙 Bingtang Orange	华宁县 Huaning county		15	MK11	华宁县 Huaning county
5	BJ2	华宁县 Huaning county		16	芽变6号 Bud sport No6.	新平县 Xinping county	4
6	SC3	华宁县 Huaning county		17	芽变7号 Bud sport No7.	新平县 Xinping county	4
7	芽变4号 Bud sport No4.	华宁县 Huaning county	不详（温州蜜柑） Unknown (Satsuma)	18	芽变8号 Bud sport No8.	新平县 Xinping county	4
8	宫本 Miyanoto Wase	华宁县 Huaning county		19	芽变9号 Bud sport No9.	新平县 Xinping county	4
9	大浦 Ooura Wase	华宁县 Huaning county		20	芽变10号 Bud Sport No10.	新平县 Xinping county	4
10	兴津 Okitsu Wase	华宁县 Huaning county		21	芽变11号 Bud sport No11.	新平县 Xinping county	4
11	日南1号 Nichihan No1.	华宁县 Huaning county		22	HG12	新平县 Xinping county

引物编号 Primer number	上游Forward (5′-3′)	下游Reward（5′-3′）
MK7	CTGGGGCTCACATAAATCGT	GATTTGTCCGCCAATCAAGT
MK9	GCATTGCAGCACTTTTGTCAT	GCATTGCAGCACTTTTGTCAT
MK19	GGGTTGGGAATGTGAATGAA	CCTAGGGGTGGGCATATTTT
MK17	CCCACCTCGTTGAATCTCTC	ACCAGTACCACTGCTTACTCTTTT

3.CHROM	3.POS	3.REF	3.ALT	7.ALT	3.DP	7.DP	3.AF	7.AF	3.SGACOV	7.SGACOV
SCAFFOLD_2	9464551	A	C		7955		.212		1688
SCAFFOLD_2	9464547	GAGAAGAAGAAGAAGA	GAGAAGAAGAAGA	GAGAAGAAGAAGA	7954	6366	.342	.397	2718	2528
SCAFFOLD_2	9464536	GAAA	GA	GA	7919	6328	.347	.409	2750	2590
SCAFFOLD_2	9464527	G	C	C	7914	6328	.366	.427	2894	2703
SCAFFOLD_2	9464521	A	G		7912		.213		1689
SCAFFOLD_6	23400577	GTATTATTATTATTATT	GTATTATTATTATTATTATTATT	GTATTATTATTATTATTATTATT	5603	5709	.606	.373	3396	2131
SCAFFOLD_6	23400577	GTATTATTATTATTATT	GTATTATTATTATTATTATTATT	T	5603	5709	.606	.227	3396	1298

利用Target SSR-seq技术鉴定温州蜜柑芽变材料

Identification of Bud Sport Mutation of Satsuma Mandarin by Target SSR-seq Technology

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