Identification of S-genotypes of 63 Pummelo Germplasm Resources

doi:10.16420/j.issn.0513-353x.2021-0260

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (5): 1111-1120.doi: 10.16420/j.issn.0513-353x.2021-0260

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Identification of S-genotypes of 63 Pummelo Germplasm Resources

WEI Zhuangmin¹^,⁴, WEI Sijia¹, CHEN Peng², HU Jianbing¹, TANG Yuqing³, YE Junli¹, LI Xianxin², DENG Xiuxin¹, CHAI Lijun¹^,^**()

¹Key Laboratory of Horticultural Plant Biology（Ministry of Education）,Huazhong Agricultural University,Wuhan 430070,China
²Institute of Horticulture,Hunan Academy of Agricultural Sciences,Changsha 410125,China
³Institute of Horticulture,Jiangxi Academy of Agricultural Sciences,Nanchang 330200,China
⁴Guangxi Subtropical Crops Research Institute,Nanning 530001,China

Received:2022-01-19 Revised:2022-02-05 Online:2022-05-25 Published:2022-05-25
Contact: CHAI Lijun E-mail:chailijun@mail.hzau.edu.cn

Abstract

Abstract:

Based on 63 pummelo germplasm resources,new S-RNases were identified by RNA-seq and sequence analysis. Twenty-one pairs of specific primers were designed to amplify and identify the S-genotypes of 63 pummelo germplasm resources. Results showed twelve new S-RNases were found,named S₁₀-RNase-S₂₁-RNase in sequence. Specific amplification showed that 59 out of 63 materials were identified with complete S-genotype,only one S-RNase gene was identified in four materials. Among them,S₂-RNase was amplified in 22 materials with the highest frequency of 34.92%（22/63）,while S₄-RNase and S₆-RNase were not amplified in the 63 materials. The results also showed that many varieties had the same S-genotype.

Key words: Citrus, S-genotypes, self-incompatibility, specific primer

CLC Number:

S666.3

WEI Zhuangmin, WEI Sijia, CHEN Peng, HU Jianbing, TANG Yuqing, YE Junli, LI Xianxin, DENG Xiuxin, CHAI Lijun. Identification of S-genotypes of 63 Pummelo Germplasm Resources[J]. Acta Horticulturae Sinica, 2022, 49(5): 1111-1120.

Figures/Tables 6

References 33

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材料 Sample	名称 Name	基因型Genotype
柚 Pummelo	马家柚Majia Pummelo	S₁₀S₁₆
	慈利甜柚3号 Cili Tian Pummelo 3	S₃S₁₁
	早熟高糖香柚3号 Early-maturing and high-sugar Xiang Pummelo 3	S₁₃S₁₄
	无核安江香柚 Wuhe Anjiang Xiang Pummelo	S₁₃S₁₄
	龙都早香柚 Longdu Early-maturing Xiang Pummelo	S₁₄S₁₅
	慈利砧板柚 Cili Zhenban Pummelo	S₁₄S₁₆
	大果红瓤柚 Big Red Pummelo	S₉S₁₇
	金兰柚 Jinlan Pummelo	S₉S₁₇
	石榴柚 Shiliu Pummelo	S₁₇
	慈利菠萝香柚2号 Cili Boluo Xiang Pummelo 2	S₂S₁₈
	慈利金香柚 Cili Jinxiang Pummelo	S₁₈S₁₉
	狮子头柚 Shizitou Pummelo	S₂S₂₁
葡萄柚 Grapefruit	胡柚（常山）Hu Pummelo（Changshan）	S₁₁S₂₀
柠檬 Lemon	香水柠檬 Xiangshui Lemon	S₁₂

材料 Sample	名称 Name	基因型Genotype
柚 Pummelo	马家柚Majia Pummelo	S₁₀S₁₆
	慈利甜柚3号 Cili Tian Pummelo 3	S₃S₁₁
	早熟高糖香柚3号 Early-maturing and high-sugar Xiang Pummelo 3	S₁₃S₁₄
	无核安江香柚 Wuhe Anjiang Xiang Pummelo	S₁₃S₁₄
	龙都早香柚 Longdu Early-maturing Xiang Pummelo	S₁₄S₁₅
	慈利砧板柚 Cili Zhenban Pummelo	S₁₄S₁₆
	大果红瓤柚 Big Red Pummelo	S₉S₁₇
	金兰柚 Jinlan Pummelo	S₉S₁₇
	石榴柚 Shiliu Pummelo	S₁₇
	慈利菠萝香柚2号 Cili Boluo Xiang Pummelo 2	S₂S₁₈
	慈利金香柚 Cili Jinxiang Pummelo	S₁₈S₁₉
	狮子头柚 Shizitou Pummelo	S₂S₂₁
葡萄柚 Grapefruit	胡柚（常山）Hu Pummelo（Changshan）	S₁₁S₂₀
柠檬 Lemon	香水柠檬 Xiangshui Lemon	S₁₂

引物名称 Primer name	引物序列（5′-3′） Sequence of primer	退火温度/℃ Annealing temperature
S₁	F：ATGAACATTACTTTCTTCCTCTACA;R：GTTGATCTCCGTTCTTCGTG	57
S₂	F：GACTAACCTCTTTCGCTTTGC;R：CGGATCCATGCCTTTTCTAG	59
S₃	F：TCCAACATCACCTATTGTACAGC;R：GATAACGACCATCACTAAAAACG	59
S₄	F：GTTTGTTTCCTGCATTTCCTC;R：GACCCCTGTTCTTTTTTACTGC	59
S₅	F：ATGAAGGTGGCATCCATCA;R：AATAATATCCGGCCCACAG	59
S₆	F：ATGGGGACTAATTTCCTCATTATCTTT;R：CTATATTTTAACGTATCGCGGCAA	59
S₇	F：ATCATCTCGGGCGCTGTTC;R：TCCCCACAGTTCTCGGTTTT	60
S₈	F：CATTACCTATTCTGCTGCTCA;R：CCGAGTAGGGAACATGATTG	59
S₉	F：GTTGTAACGCAAAACACTTCTG;R：CGTATGAGCATGTTAGTCTTGG	59
S₁₀	F：CTGCATTGCCTCCGGCAC;R：TCGCCATTTTAACGGATCGAGG	60
S₁₁	F：GCCGCTCCAAACTCTTTCG;R：CGCAGGGAACTTGATGTTGT	60
S₁₂	F：TGCTGCAATGGCCAACTCTT;R：CCGGGTGGGGAACTTGATG	60
S₁₃	F：TCCACCACTTTTGGCTGGTC;R：TCTGCGCAACATTCGTCTCA	61
S₁₄	F：AACGAAGGCAGCTTACCTCT;R：CACGGCAGCTCTCTCCATTA	59
S₁₅	F：CAGCCTTGCCTCGTTTGAAG;R：GCTTGATCGTCAGCGCAAAG	59
S₁₆	F：CTACATCATAAATTCTTCCGGGC;R：GTGGTTTTGGTCGTTTTCCC	61
S₁₇	F：GCTCGGGGTTCCAGAAAGT;R：ACGGTTCAGGAAACTTGATACG	61
S₁₈	F：GCAGCTCAAAACTCGTCGC;R：TAAACGGCAGGGGGAGAGAA	59
S₁₉	F：GCTCAAGCTGACCACTGTTG;R：CTACCAGCGTGGAGGGAAAATA	59
S₂₀	F：TCAGGTGCAGCAGAAAACAC;R：CTTGGCCGTCCACGCATATAA	59
S₂₁	F：TCCACCACTTTTGGCTGGTC;R：CGGGTTTGGTTCGGGAAACT	59

引物名称 Primer name	引物序列（5′-3′） Sequence of primer	退火温度/℃ Annealing temperature
S₁	F：ATGAACATTACTTTCTTCCTCTACA;R：GTTGATCTCCGTTCTTCGTG	57
S₂	F：GACTAACCTCTTTCGCTTTGC;R：CGGATCCATGCCTTTTCTAG	59
S₃	F：TCCAACATCACCTATTGTACAGC;R：GATAACGACCATCACTAAAAACG	59
S₄	F：GTTTGTTTCCTGCATTTCCTC;R：GACCCCTGTTCTTTTTTACTGC	59
S₅	F：ATGAAGGTGGCATCCATCA;R：AATAATATCCGGCCCACAG	59
S₆	F：ATGGGGACTAATTTCCTCATTATCTTT;R：CTATATTTTAACGTATCGCGGCAA	59
S₇	F：ATCATCTCGGGCGCTGTTC;R：TCCCCACAGTTCTCGGTTTT	60
S₈	F：CATTACCTATTCTGCTGCTCA;R：CCGAGTAGGGAACATGATTG	59
S₉	F：GTTGTAACGCAAAACACTTCTG;R：CGTATGAGCATGTTAGTCTTGG	59
S₁₀	F：CTGCATTGCCTCCGGCAC;R：TCGCCATTTTAACGGATCGAGG	60
S₁₁	F：GCCGCTCCAAACTCTTTCG;R：CGCAGGGAACTTGATGTTGT	60
S₁₂	F：TGCTGCAATGGCCAACTCTT;R：CCGGGTGGGGAACTTGATG	60
S₁₃	F：TCCACCACTTTTGGCTGGTC;R：TCTGCGCAACATTCGTCTCA	61
S₁₄	F：AACGAAGGCAGCTTACCTCT;R：CACGGCAGCTCTCTCCATTA	59
S₁₅	F：CAGCCTTGCCTCGTTTGAAG;R：GCTTGATCGTCAGCGCAAAG	59
S₁₆	F：CTACATCATAAATTCTTCCGGGC;R：GTGGTTTTGGTCGTTTTCCC	61
S₁₇	F：GCTCGGGGTTCCAGAAAGT;R：ACGGTTCAGGAAACTTGATACG	61
S₁₈	F：GCAGCTCAAAACTCGTCGC;R：TAAACGGCAGGGGGAGAGAA	59
S₁₉	F：GCTCAAGCTGACCACTGTTG;R：CTACCAGCGTGGAGGGAAAATA	59
S₂₀	F：TCAGGTGCAGCAGAAAACAC;R：CTTGGCCGTCCACGCATATAA	59
S₂₁	F：TCCACCACTTTTGGCTGGTC;R：CGGGTTTGGTTCGGGAAACT	59

材料 Sample	序号 No.	名称 Name	采集地 Origin	基因型 Genotype
柚	1	金沙柚6号 Jinsha Pummelo 6	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₂S₃
Pummelo	2	金沙柚4号 Jinsha Pummelo 4	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₁Sx
	3	金沙柚2号 Jinsha Pummelo 2	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₂S₃
	4	金沙柚1号 Jinsha Pummelo 1	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₁S₅
	5	沙田柚（南昌）Shatian Pummelo（Nanchang）	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₁S₂
	6	少核沙田柚 Shaohe Shatian Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₁S₂
	7	无核沙田柚 Wuhe Shatian Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₁S₂
	10	江坝柚 Jiangba Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₂S₂₁
	11	丰城红壤柚 Fengcheng Red Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₂S₃
	12	谢家柚 Xiejia Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₂S₃
	13	蔡家柚 Caijia Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₁₁S₁₂
	14	美国橙柚 America Orange-Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₁₃S_x
	16	鹅旦柚 Edan Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₂S₃
	17	狮子头柚 Shizitou Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₂S₂₁
	18	红壤文旦柚 Hongrang Wendan Pummelo	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₂S₃
	19	晚白柚 Wanbai Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₂S₅
	20	菊花芯柚 Juhuaxin Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₃S₁₁
	21	早熟高糖香柚4号 Early-maturing and high-sugar Xiang Pummelo 4	湖南省怀化市 Huaihua,Hunan Province	S₈S₁₅
	22	金兰柚 Jinlan Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₉S₁₇
	23	三蔸半柚 Sandouban Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₁₂S₁₃
	24	慈利香柚 Cili Xiang Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₁S₂
	25	文旦柚 Wendan Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₈S₁₇
	26	本地变异柚 Native Mutational Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₉S₁₇
	27	牛腿柚 Niutui Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₁₁S₂₁
	28	张家界本地红心柚 Zhangjiajie Native Red Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₁₀S₁₆
	29	强德勒红心柚 Red Chandler	湖南省农业科学院 Hunan Academy of Agricultural Sciences	S₉S₁₇
	30	慈利甜柚3号 Cili Tian Pummelo 3	湖南省张家界市 Zhangjiajie,Hunan Province	S₃S₁₁
	31	糯米柚 Nuomi Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₁₁S₁₄
	32	沙田柚-1（长沙） Shatian Pummelo 1（Changsha）	湖南省怀化市 Huaihua,Hunan Province	S₁S₂
	33	早熟高糖香柚3号 Early-maturing and high-sugar Xiangyou 3	湖南省怀化市 Huaihua,Hunan Province	S₁₃S₁₄
	34	冰糖柚 Bingtang Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₅S₁₃
	35	中秋蜜柚 Zhongqiu Mi Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₈S₉
	36	早熟高糖香柚1号 Early-maturing and high-sugar Xiangyou 1	湖南省怀化市 Huaihua,Hunan Province	S₈S₁₅
	37	永嘉早香柚 Yongjia Early-maturing xiang Pummelo	浙江省温州市 Wenzhou,Zhejiang Province	S₂S₈
	38	早熟高糖香柚2号 Early-maturing and high-sugar Xiangyou 2	湖南省怀化市 Huaihua,Hunan Province	S₈S₁₇
	39	慈利菠萝香柚2号Cili Boluo Xiang Pummelo 2	湖南省张家界市 Zhangjiajie,Hunan Province	S₂S₁₈
	40	安农1号 Annong Pummelo 1	湖南省怀化市 Huaihua,Hunan Province	S₁₃S₁₄
	41	安江无核红心柚母株 Mother plant of Anjiang Wuhe Red Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₉S₁₇
	42	白玉霜柚 Baiyushuang Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₁₄S_x
	43	大果红瓤柚 Big Red Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₉S₁₇
	44	假石榴柚 Jiashiliu Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₁₀S₁₃
	45	无核安江香柚Wuhe Anjiang Xiang Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₁₃S₁₄
	46	安江无核红心柚子代 Offspring of Anjiang Wuhe Hongxinyou	湖南省怀化市 Huaihua,Hunan Province	S₉S₁₇
材料 Sample	序号 No.	名称 Name	采集地 Origin	基因型 Genotype
	47	慈利矮晚柚 Cili Aiwan Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₂S₅
	48	安江红心柚 Anjiang Red Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₁₀S₁₆
	49	马家柚 Majia Pummelo	江西省上饶市 Shangrao,Jiangxi Province	S₁₀S₁₆
	50	安江酸柚 Anjiang Sour Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₂S₅
	51	慈利金香柚 Cili Jinxiang Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₁₈S₁₉
	52	龙都早香柚 Longdu Early-maturing Xiang Pummelo	四川省自贡市 Zigong,Sichuan Province	S₁₄S₁₅
	53	慈利甜柚1号 Cili Tian Pummelo 1	湖南省张家界市 Zhangjiajie,Hunan Province	S₁S₇
	54	慈利砧板柚 Cili Zhenban Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₁₄S₁₆
	55	沙田柚-2（长沙）Shatian Pummelo 2（Changsha）	湖南省怀化市 Huaihua,Hunan Province	S₁S₂
	56	安江香柚 Anjiang Xiang Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₉S₁₃
	57	石榴柚 Shiliu Pummelo	湖南省怀化市 Huaihua,Hunan Province	S₁₇S_x
	58	慈利菠萝香柚1号Cili Boluo Xiang Pummelo 1	湖南省张家界市 Zhangjiajie,Hunan Province	S₂S₁₈
	59	空心塔柚 Kongxinta Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₉S₁₇
	60	永定白柚 Yongding White Pummelo	湖南省张家界市 Zhangjiajie,Hunan Province	S₅S₁₀
	61	未知柚1号 Unknown Pummelo 1	湖南省农业科学院 Hunan Academy of Agricultural Sciences	S₁₃S₁₄
	62	未知柚2号 Unknown Pummelo 2	湖南省农业科学院 Hunan Academy of Agricultural Sciences	S₂S₂₁
	63	未知柚3号 Unknown Pummelo 3	湖南省怀化市 Huaihua,Hunan Province	S₉S₁₇
葡萄柚	8	胡柚（衢州）Hu Pummelo（Quzhou）	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₁₁S₂₀
Grapefruit	9	胡柚（常山）Hu Pummelo（Changshan）	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₁₁S₂₀
	15	葡萄柚Grapefruit	江西省农业科学院 Jiangxi Academy of Agricultural Sciences	S₂S₃

Identification of S-genotypes of 63 Pummelo Germplasm Resources

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