利用PCR产物混池重测序检测梨褐皮关联SLAF 标签多态性

doi:10.16420/j.issn.0513-353x.2023-0263

摘要/Abstract

摘要：

在已知梨褐皮关联的SLAF标签（Marker286117）序列上存在4个SNP位点，共有4个基因型（a ~ d）的基础上，使用PCR加混池重测序技术鉴定该SLAF标签在64个梨品种中的多态性。共获得7.1 Gb原始数据，通过编译的SNP-rseq和Calculate程序对数据进行处理，64个梨样品获得的平均read数量为117 642条，满足分析需求。在该SLAF标签上新发现2个基因型（e和f）。6个基因型中c为原始类型，在栽培种砂梨和白梨中鉴定到b、d和e基因型；a和f基因型则出现在拥有西方梨血统的样品中。对基因型和表型统计后发现，c基因型存在时，梨果皮为褐色，不含c基因型时为绿色。该规律不仅适用于东方梨，在西方梨的一些样品中也适用。混池重测序技术降低了测序成本，相关技术手段能够应用于简化重测序后续的验证和标记辅助育种。

关键词: 梨, SLAF标签, SNP, 多态性, 重测序

Abstract:

Previous studies have shown that there are four SNP loci on the sequence of the russet peel associated SLAF tag（Marker286117），with a total of four genotypes（a，b，c，d） in Pyrus. In this study，PCR plus mixing pool re-sequencing was used to identify the polymorphisms of the SLAF tag in 64 pear cultivars. The results showed that a total of 7.1 Gb of raw data was obtained by re-sequencing，and the data were processed by two programs SNP-rseq and Calculate. The average number of reads obtained from 64 pear accessions was 117 642，which met the analysis needs. Two new genotypes（e and f） were found on this SLAF locus. Among six genotypes，genotype c was the original type，and genotypes b，d and e were produced in cultivated pears of P. pyrifolia and P. bretschneideri. Genotypes a and f appeared in samples of Western pear. When genotype and phenotype statistics showed that only genotype c existed，the peel of the pear fruit was russet，and those without genotype c were green. This rule applies not only to Eastern pears，but also to some Western pears. In this study，mixed-pool was used to reduce the cost of sequencing. This method could be applied to simplify the subsequent validation of re-sequencing and marking-assisted breeding.

Key words: Pyrus, SLAF tag, SNP, polymorphism, re-sequencing

蒋爽, 王晓庆, 施春晖, 李水根, 骆军. 利用PCR产物混池重测序检测梨褐皮关联SLAF 标签多态性[J]. 园艺学报, 2024, 51(4): 707-714.

JIANG Shuang, WANG Xiaoqing, SHI Chunhui, LI Shuigen, LUO Jun. Detection of SLAF Tag Polymorphism Associated with Pear Russet Peel by PCR Product Pool Resequencing[J]. Acta Horticulturae Sinica, 2024, 51(4): 707-714.

图/表 4

参考文献 18

[1]	Doyle J J, Doyle J L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull, 19:11-15.
[2]	Fan Tao, Li Zhi, Jiang Qing, Chen Shulin, Ou Xia, Chen Yongyan. 2021. Development and effect evaluation of KASP markers closely linked to major QTLs of spike number per unit area and grain length in wheat. Scientia Agricultura Sinica, 54 (14):11. (in Chinese)
	范涛, 李治, 蒋庆, 陈姝霖, 欧霞, 陈永艳. 2021. 小麦单位面积穗数和粒长主效QTL紧密连锁KASP标记的开发及其效应评价. 中国农业科学, 54 (14):11.
[3]	Geng X, Jiang C, Yang J, Wang L, Wu X, Wei W. 2016. Rapid identification of candidate genes for seed weight using the SLAF-Seq method in Brassica napus. PLoS ONE, 11 (1):e0147580.
[4]	Jiang S, Luo J, Wang X Q, An H S, Zhang J Y, Li S G. 2022. QTL mapping and transcriptome analysis to identify genes associated with green/russet peel in Pyrus pyrifolia. Scientia Horticulturae, 293:110714.
[5]	Khera P, Upadhyaya H D, Pandey M K, Roorkiwal M, Sriswathi M, Janila P, Guo Y, McKain M R, Nagy E D, Knapp S J, Leebens-Mack J, Conner J A, Ozias-Akins P, Varshney R K. 2013. Single nucleotide polymorphism-based genetic diversity in the reference set of peanut (Arachis spp.) by developing and applying cost-effective kompetitive allele specific polymerase chain reaction genotyping assays. The Plant Genome, 6 (3):0019.
[6]	Kumar S, Kirk C, Deng C, Wiedow C, Knaebel M, Brewer L. 2017. Genotyping-by-sequencing of pear(Pyrus spp.)accessions unravels novel patterns of genetic diversity and selection footprints. Horticulture Research, 4:17015.
[7]	Lian Yun, Li Hai-chao, Li Jin-ying. 2021. Marker-assisted screening of soybean cyst nematode germplasms harboring resistance loci rhg1 and Rhg4. Journal of Plant Genetic Resources, 22 (2):399-406. (in Chinese)
	练云, 李海朝, 李金英. 2021. 利用KASP标记筛选含rhg1和Rhg4位点的大豆抗病资源. 植物遗传资源学报, 22 (2):399-406.
[8]	Meng Junren, Ceng Wenfang, Deng Li, Pan Lei, Lu Zhenhua, Cui Guochao, Wang Zhiqiang, Niu Liang. 2021. Development and application of KASP molecular markers of some important traits for peach. Scientia Agricultura Sinica,(15):3295-3307. (in Chinese) doi: 10.3864/j.issn.0578-1752.2021.15.013
	孟君仁, 曾文芳, 邓丽, 潘磊, 鲁振华, 崔国朝, 王志强, 牛良. 2021. 桃若干重要性状的KASP分子标记开发与应用. 中国农业科学,(15):3295-3307. doi: 10.3864/j.issn.0578-1752.2021.15.013
[9]	Ren Hailong, Xu Donglin, Zhang Jing, Zou Jiwen, Li Guangguang, Zhou Xianyu, Xiao Wanyu, Sun Yijia. 2023. Establishment of SNP fingerprinting and identification of Chinese flowering cabbage varieties based on KASP genotyping. Acta Horticulturae Sinica, 50 (2):307-318. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-1046
	任海龙, 许东林, 张晶, 邹集文, 李光光, 周贤玉, 肖婉钰, 孙艺嘉. 2023. 菜薹KASP-SNP指纹图谱构建及品种鉴定. 园艺学报, 50 (2):307-318. doi: 10.16420/j.issn.0513-353x.2021-1046
[10]	Shi P, Xu Z, Zhang S Y, Wang X J, Ma X F, Zheng J C, Xing L B, Zhang D, Ma J J, Han M Y, Zhao C P. 2020. Construction of a high-density SNP-based genetic map and identification of fruit-related QTLs and candidate genes in peach [Prunus persica (L.) Batsch]. BMC Plant Biology, 20 (1):438.
[11]	Singh N, Choudhury D R, Singh A K, Kumar S, Srinivasan K, Tyagi R K, Singh N K, Singh R. 2013. Comparison of SSR and SNP markers in estimation of genetic diversity and population structure of Indian rice varieties. PLoS ONE, 8 (12):e84136.
[12]	Sun X, Liu D, Zhang X, Li W, Liu H, Hong W, Jiang C, Ning G, Ma C, Zeng H. 2013. SLAF-seq:an efficient method of large-scale de novo SNP discovery and genotyping using high-throughput sequencing. PLoS ONE, 8 (3):e58700.
[13]	Wang L, Li X, Wang L, Xue H, Wu J, Yin H, Zhang S. 2017. Construction of a high-density genetic linkage map in pear(Pyrus communis × Pyrus pyrifolia Nakai)using SSRs and SNPs developed by SLAF-seq. Scientia Horticulturae, 218:198-204.
[14]	Wu J, Li L T, Li M, Khan M A, Li X G, Chen H, Yin H, Zhang S L. 2014. High-density genetic linkage map construction and identification of fruit-related QTLs in pear using SNP and SSR markers. Journal of Experimental Botany, 65 (20):5771-5781. doi: 10.1093/jxb/eru311 pmid: 25129128
[15]	Wu Zhijun, Li Wei, Wang Xinghua, Zhang Cheng, Jiang Xun, Li Sheng, Luo Liyong, Sun Kang, Zeng Liang. 2022. Genetic relationship analysis of the population germplasms of section Thea with unstable ovary locule number based on SLAF-seq. Acta Horticulturae Sinica, 49 (11):2455-2470. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0876
	吴致君, 李伟, 王兴华, 张成, 蒋勋, 黎盛, 罗理勇, 孙康, 曾亮. 2022. 基于SLAF-seq的茶组子房室数不稳定居群种质亲缘关系分析. 园艺学报, 49 (11):2455-2470. doi: 10.16420/j.issn.0513-353x.2021-0876
[16]	Xu X, Xu R, Zhu B, Yu T, Qu W, Lu L, Xu Q, Qi X, Chen X. 2014. A high-density genetic map of cucumber derived from Specific Length Amplified Fragment sequencing(SLAF-seq). Frontiers in Plant Science, 5:768.
[17]	Yang J J, Zhang J, Du H S, Zhao H, Mao A J, Zhang X F, Jiang L, Zhang H Y, Wen C L, Xu Y. 2022. Genetic relationship and pedigree of Chinese watermelon varieties based on diversity of perfect SNPs. Horticultural Plant Journal, 8 (4):489-498.
[18]	Zhang J, Yang J J, Fu S Z, Ren J, Zhang X F, Xia C X, Zhao H, Yang K, Wen C L. 2022. Comparison of DUS testing and SNP fingerprinting for variety identification in cucumber. Horticultural Plant Journal, 8 (5):575-582.

上游引物；下游引物 Forward primer；reverse primer	编号 ID	样品名称 Sample name	种 Species	果皮 Peel	检出条带数 Number of reads	基因型 Genotype
AGAGGA_MF；MR	1	J188	砂梨Pyrus pyrifolia	褐Russet	116 146	ce
CTAAGA_MF；MR	2	沪晓Huxiao	砂梨P. pyrifolia	褐Russet	83 486	cd
TCCGGT_MF；MR	3	沪晶梨18号Hujingli 18	砂梨P. pyrifolia	褐Russet	35 355	ce
GGGAGT_MF；MR	4	沪晶梨67号Hujingli 67	砂梨P. pyrifolia	褐Russet	68 550	bc
GTCCTC_MF；MR	5	翠冠Cuiguan	砂梨P. pyrifolia	绿^*Green	160 234	be
CGCTTA_MF；MR	6	苏翠1号Sucui 1	砂梨P. pyrifolia	绿Green	18 072	bb
GGCTGG_MF；MR	7	翠玉Cuiyu	砂梨P. pyrifolia	绿Green	41 657	bb
TTGGTC_MF；MR	8	鸭梨Yali	白梨P. bretschneideri	绿Green	48 436	bb
GACCCT_MF；MR	9	珍珠梨Zhenzhuli	砂梨 × 西洋梨 P. Pyrifolia × P. communis	绿Green	111 934	bf
TACAGG_MF；MR	10	黄金梨Whangkeumbae	砂梨P. pyrifolia	绿Green	15 917	bb
AGTTCC_MF；MR	11	金二十世纪Gold Nijisseiki	砂梨P. pyrifolia	绿Green	84 980	bd
GGCGCC_MF；MR	12	黄花Huanghua	砂梨P. pyrifolia	褐Russet	105 373	ce
AAACTC_MF；MR	13	筑水Chikusui	砂梨P. pyrifolia	褐Russet	88 559	bc
TTTCCC_MF；MR	14	八幸Hakko	砂梨P. pyrifolia	绿Green	44 236	be
GGACGC_MF；MR	15	丰水Housui	砂梨P. pyrifolia	褐Russet	65 746	cc
CCCTAC_MF；MR	16	圆黄Wonhwang	砂梨P. pyrifolia	褐Russet	84 151	cc
TAGACA_MF；MR	17	黄冠Huangguan	砂梨P. pyrifolia	绿Green	63 316	bb
GTAAAG_MF；MR	18	雪青Xueqing	砂梨P. pyrifolia	绿Green	102 593	bb
GATCAA_MF；MR	19	香梨Xiangli	砂梨P. pyrifolia	绿Green	93 861	de
CAGGGT_MF；MR	20	菊水Kikusui	砂梨P. pyrifolia	绿Green	98 341	de
ACAAGG_MF；MR	21	满丰Mansoo	砂梨P. pyrifolia	褐Russet	125 950	bc
CCCGCG_MF；MR	22	新高Niitaka	砂梨P. pyrifolia	褐Russet	76 014	cd
GCGCGA_MF；MR	23	新水Shinsui	砂梨P. pyrifolia	褐Russet	69 427	cd
CTTTGT_MF；MR	24	新世纪Shinseki	砂梨P. pyrifolia	绿Green	47 533	bb
CGGTAG_MF；MR	25	早生新水Zaosheng Shinsui	砂梨P. pyrifolia	褐Russet	145 075	cd
ACACCT_MF；MR	26	秋水Qiushui	砂梨P. pyrifolia	绿Green	168 616	bb
CGTAAA_MF；MR	27	川梨1 P. pashia 1	川梨P. pashia	褐Russet	29 657	cc
GCCGAA_MF；MR	28	豆梨1 P. calleryana 1	豆梨P. calleryana	褐Russet	38 034	cc
GGATCA_MF；MR	29	杜梨1 P. betulifolia 1	杜梨P. betulifolia	褐Russet	93 720	cc
ATCGCA_MF；MR	30	考密斯Comice	西洋梨P. communis	绿Green	98 436	af
TCGTCG_MF；MR	31	雪梨 P. nivalis	雪梨P. nivalis	绿Green	34 630	ff
GGTATC_MF；MR	32	柳叶梨 P. salicifolia	柳叶梨P. salicifolia	褐Russet	34 577	ce
AGAGGA_MF；MR	33	八达Bada	砂梨P. pyrifolia	绿Green	305 720	bb
CTAAGA_MF；MR	34	铁头Tietou	砂梨 × 西洋梨 P. Pyrifolia × P. communis	褐Russet	121 735	cf
TCCGGT_MF；MR	34	爱宕Atago	砂梨P. pyrifolia	褐Russet	89 989	bc
GGGAGT_MF；MR	36	YASTE	砂梨P. pyrifolia	绿Green	74 786	be
GTCCTC_MF；MR	37	极光Jiguang	西洋梨P. communis	绿Green	112 567	ff
CGCTTA_MF；MR	38	晚三吉Okusankichi	砂梨P. pyrifolia	褐Russet	51 847	cc
GGCTGG_MF；MR	39	苍溪梨Cangxi	砂梨P. pyrifolia	褐Russet	37 581	bc
TTGGTC_MF；MR	40	八里香Balixiang	秋子梨P.ussuriensis	绿Green	103 489	bb
GACCCT_MF；MR	41	康德Kande	砂梨 × 西洋梨 P. Pyrifolia × P. communis	绿Green	119 982	bf
TACAGG_MF；MR	42	鸭蛋青Yadangqing	砂梨P. pyrifolia	绿Green	5 384	bb
AGTTCC_MF；MR	43	满天红Mantianhong	砂梨P. pyrifolia	绿Green	896 836	be
GGCGCC_MF；MR	44	巴梨Bartlett	西洋梨P. communis	绿Green	81 781	af
AAACTC_MF；MR	45	华山Huashan	砂梨P. pyrifolia	褐Russet	304	bc
TTTCCC_MF；MR	46	今村秋Imanuraaki	砂梨P. pyrifolia	褐Russet	236 954	cc
GGACGC_MF；MR	47	La France	西洋梨P. communis	绿Green	138 556	aa
CCCTAC_MF；MR	48	新酥梨Xinsu	砂梨P. pyrifolia	绿Green	268 804	bb
TAGACA_MF；MR	49	雁荡雪梨Yandang Xueli	砂梨P. pyrifolia	绿Green	373 778	bb
GTAAAG_MF；MR	50	早酥Zaosu	砂梨P. pyrifolia	绿Green	123 279	bf
GATCAA_MF；MR	51	大冬果Dadongguo	砂梨P. pyrifolia	绿Green	109 604	bb
CAGGGT_MF；MR	52	黄皮钟Huangpizhong	砂梨P. pyrifolia	褐Russet	197 363	cc
ACAAGG_MF；MR	53	满顶雪梨Manding Xueli	砂梨P. pyrifolia	绿Green	166 382	bc
CCCGCG_MF；MR	54	伊春山梨Yichun Shanli	秋子梨P.ussuriensis	褐Russet	28 740	bc
GCGCGA_MF；MR	55	四平山梨Siping Shanli	秋子梨P.ussuriensis	褐Russet	18 712	be
CTTTGT_MF；MR	56	杜梨2 P. betulifolia 2	杜梨P. betulifolia	褐Russet	88 082	cc
CGGTAG_MF；MR	57	川梨2 P. pashia 2	川梨P. pashia	褐Russet	233 002	cc
ACACCT_MF；MR	58	豆梨2 P. calleryana 2	豆梨P. calleryana	褐Russet	251 969	cc
CGTAAA_MF；MR	59	秋荣Qiuron	砂梨P. pyrifolia	褐Russet	93 051	cd
GCCGAA_MF；MR	60	砀山酥梨Dangshan Suli	白梨P. bretschneideri	绿Green	36 319	bb
GGATCA_MF；MR	61	Hougesi	砂梨P. pyrifolia	褐Russet	135 358	bc
ATCGCA_MF；MR	62	阿巴特Abate Fetel	西洋梨P. communis	绿Green	320 099	ff
TCGTCG_MF；MR	63	南果梨Nanguoli	秋子梨P.ussuriensis	绿Green	25 869	bb
GGTATC_MF；MR	64	花盖梨Huagaili	秋子梨P.ussuriensis	褐Russet	188 592	bc

上游引物；下游引物 Forward primer；reverse primer	编号 ID	样品名称 Sample name	种 Species	果皮 Peel	检出条带数 Number of reads	基因型 Genotype
AGAGGA_MF；MR	1	J188	砂梨Pyrus pyrifolia	褐Russet	116 146	ce
CTAAGA_MF；MR	2	沪晓Huxiao	砂梨P. pyrifolia	褐Russet	83 486	cd
TCCGGT_MF；MR	3	沪晶梨18号Hujingli 18	砂梨P. pyrifolia	褐Russet	35 355	ce
GGGAGT_MF；MR	4	沪晶梨67号Hujingli 67	砂梨P. pyrifolia	褐Russet	68 550	bc
GTCCTC_MF；MR	5	翠冠Cuiguan	砂梨P. pyrifolia	绿^*Green	160 234	be
CGCTTA_MF；MR	6	苏翠1号Sucui 1	砂梨P. pyrifolia	绿Green	18 072	bb
GGCTGG_MF；MR	7	翠玉Cuiyu	砂梨P. pyrifolia	绿Green	41 657	bb
TTGGTC_MF；MR	8	鸭梨Yali	白梨P. bretschneideri	绿Green	48 436	bb
GACCCT_MF；MR	9	珍珠梨Zhenzhuli	砂梨 × 西洋梨 P. Pyrifolia × P. communis	绿Green	111 934	bf
TACAGG_MF；MR	10	黄金梨Whangkeumbae	砂梨P. pyrifolia	绿Green	15 917	bb
AGTTCC_MF；MR	11	金二十世纪Gold Nijisseiki	砂梨P. pyrifolia	绿Green	84 980	bd
GGCGCC_MF；MR	12	黄花Huanghua	砂梨P. pyrifolia	褐Russet	105 373	ce
AAACTC_MF；MR	13	筑水Chikusui	砂梨P. pyrifolia	褐Russet	88 559	bc
TTTCCC_MF；MR	14	八幸Hakko	砂梨P. pyrifolia	绿Green	44 236	be
GGACGC_MF；MR	15	丰水Housui	砂梨P. pyrifolia	褐Russet	65 746	cc
CCCTAC_MF；MR	16	圆黄Wonhwang	砂梨P. pyrifolia	褐Russet	84 151	cc
TAGACA_MF；MR	17	黄冠Huangguan	砂梨P. pyrifolia	绿Green	63 316	bb
GTAAAG_MF；MR	18	雪青Xueqing	砂梨P. pyrifolia	绿Green	102 593	bb
GATCAA_MF；MR	19	香梨Xiangli	砂梨P. pyrifolia	绿Green	93 861	de
CAGGGT_MF；MR	20	菊水Kikusui	砂梨P. pyrifolia	绿Green	98 341	de
ACAAGG_MF；MR	21	满丰Mansoo	砂梨P. pyrifolia	褐Russet	125 950	bc
CCCGCG_MF；MR	22	新高Niitaka	砂梨P. pyrifolia	褐Russet	76 014	cd
GCGCGA_MF；MR	23	新水Shinsui	砂梨P. pyrifolia	褐Russet	69 427	cd
CTTTGT_MF；MR	24	新世纪Shinseki	砂梨P. pyrifolia	绿Green	47 533	bb
CGGTAG_MF；MR	25	早生新水Zaosheng Shinsui	砂梨P. pyrifolia	褐Russet	145 075	cd
ACACCT_MF；MR	26	秋水Qiushui	砂梨P. pyrifolia	绿Green	168 616	bb
CGTAAA_MF；MR	27	川梨1 P. pashia 1	川梨P. pashia	褐Russet	29 657	cc
GCCGAA_MF；MR	28	豆梨1 P. calleryana 1	豆梨P. calleryana	褐Russet	38 034	cc
GGATCA_MF；MR	29	杜梨1 P. betulifolia 1	杜梨P. betulifolia	褐Russet	93 720	cc
ATCGCA_MF；MR	30	考密斯Comice	西洋梨P. communis	绿Green	98 436	af
TCGTCG_MF；MR	31	雪梨 P. nivalis	雪梨P. nivalis	绿Green	34 630	ff
GGTATC_MF；MR	32	柳叶梨 P. salicifolia	柳叶梨P. salicifolia	褐Russet	34 577	ce
AGAGGA_MF；MR	33	八达Bada	砂梨P. pyrifolia	绿Green	305 720	bb
CTAAGA_MF；MR	34	铁头Tietou	砂梨 × 西洋梨 P. Pyrifolia × P. communis	褐Russet	121 735	cf
TCCGGT_MF；MR	34	爱宕Atago	砂梨P. pyrifolia	褐Russet	89 989	bc
GGGAGT_MF；MR	36	YASTE	砂梨P. pyrifolia	绿Green	74 786	be
GTCCTC_MF；MR	37	极光Jiguang	西洋梨P. communis	绿Green	112 567	ff
CGCTTA_MF；MR	38	晚三吉Okusankichi	砂梨P. pyrifolia	褐Russet	51 847	cc
GGCTGG_MF；MR	39	苍溪梨Cangxi	砂梨P. pyrifolia	褐Russet	37 581	bc
TTGGTC_MF；MR	40	八里香Balixiang	秋子梨P.ussuriensis	绿Green	103 489	bb
GACCCT_MF；MR	41	康德Kande	砂梨 × 西洋梨 P. Pyrifolia × P. communis	绿Green	119 982	bf
TACAGG_MF；MR	42	鸭蛋青Yadangqing	砂梨P. pyrifolia	绿Green	5 384	bb
AGTTCC_MF；MR	43	满天红Mantianhong	砂梨P. pyrifolia	绿Green	896 836	be
GGCGCC_MF；MR	44	巴梨Bartlett	西洋梨P. communis	绿Green	81 781	af
AAACTC_MF；MR	45	华山Huashan	砂梨P. pyrifolia	褐Russet	304	bc
TTTCCC_MF；MR	46	今村秋Imanuraaki	砂梨P. pyrifolia	褐Russet	236 954	cc
GGACGC_MF；MR	47	La France	西洋梨P. communis	绿Green	138 556	aa
CCCTAC_MF；MR	48	新酥梨Xinsu	砂梨P. pyrifolia	绿Green	268 804	bb
TAGACA_MF；MR	49	雁荡雪梨Yandang Xueli	砂梨P. pyrifolia	绿Green	373 778	bb
GTAAAG_MF；MR	50	早酥Zaosu	砂梨P. pyrifolia	绿Green	123 279	bf
GATCAA_MF；MR	51	大冬果Dadongguo	砂梨P. pyrifolia	绿Green	109 604	bb
CAGGGT_MF；MR	52	黄皮钟Huangpizhong	砂梨P. pyrifolia	褐Russet	197 363	cc
ACAAGG_MF；MR	53	满顶雪梨Manding Xueli	砂梨P. pyrifolia	绿Green	166 382	bc
CCCGCG_MF；MR	54	伊春山梨Yichun Shanli	秋子梨P.ussuriensis	褐Russet	28 740	bc
GCGCGA_MF；MR	55	四平山梨Siping Shanli	秋子梨P.ussuriensis	褐Russet	18 712	be
CTTTGT_MF；MR	56	杜梨2 P. betulifolia 2	杜梨P. betulifolia	褐Russet	88 082	cc
CGGTAG_MF；MR	57	川梨2 P. pashia 2	川梨P. pashia	褐Russet	233 002	cc
ACACCT_MF；MR	58	豆梨2 P. calleryana 2	豆梨P. calleryana	褐Russet	251 969	cc
CGTAAA_MF；MR	59	秋荣Qiuron	砂梨P. pyrifolia	褐Russet	93 051	cd
GCCGAA_MF；MR	60	砀山酥梨Dangshan Suli	白梨P. bretschneideri	绿Green	36 319	bb
GGATCA_MF；MR	61	Hougesi	砂梨P. pyrifolia	褐Russet	135 358	bc
ATCGCA_MF；MR	62	阿巴特Abate Fetel	西洋梨P. communis	绿Green	320 099	ff
TCGTCG_MF；MR	63	南果梨Nanguoli	秋子梨P.ussuriensis	绿Green	25 869	bb
GGTATC_MF；MR	64	花盖梨Huagaili	秋子梨P.ussuriensis	褐Russet	188 592	bc