Identification of S Genes and SFB4' in Cerasus avium L.
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摘要:
目的 甜樱桃（Cerasus avium L.）的S基因决定其自交的亲和性。为鉴定四川地区甜樱桃品种的S基因型和高自交亲和性品种，本研究对收集到的39份甜樱桃材料进行S基因型鉴定，并比较SFB4与其自交亲和突变体SFB4'基因的差异。 方法 利用S基因通用引物和特异引物对四川地区种植的39份甜樱桃叶片DNA进行PCR扩增，并对含有SFB4/SFB4'基因的扩增片段进行测序。 结果 39份甜樱桃材料共鉴定出S1、S2、S3、S4、S6、S9等6个基因型，其中27份材料含有S3基因，19份材料含有S9基因，12份材料含有S1基因，11份材料含有S4基因，7份材料含有S6基因，3份材料含有S2基因。根据鉴定得到的S基因型结果，扩增得到含有S4基因的11份材料中的SFB基因的ORF序列，比对结果显示，相比于SFB4，SFB4'在903 bp处存在4个碱基缺失；本研究设计了SFB4'特异引物，测序结果显示，有7份材料的SFB基因型为SFB4'。 结论 共鉴定得到S1、S2、S3、S4、S6、S9等6个基因型，对含有S4基因的11份材料的S4基因全长进行测序，确定7份材料为突变基因SFB4'。本研究为四川地区甜樱桃授粉树配置提供了数据支撑，并为甜樱桃新品种的选育奠定了理论基础。 Abstract:Objective Varieties of sweet cherry cultivars from Sichuan Province were analyzed for self-compatibility by the presence of mutant S gene. Method Using the universal and specific primers of S genotype, DNA from the leaves of 39 Cerasus avium L. specimens collected in Sichuan were amplified by PCR. The amplified fragments containing SFB4 and/or SFB4' were sequenced for comparison and further analysis. Result Six genes of S genotype were identified from the 39 sweet cherry specimens with 27 of them having S3 genes, 19 having S9, 12 having S1, 11 having S4, 7 having S6, and 3 having S2. In the 11 specimens containing S4 genes, the ORF sequences of SFBs were amplified. By comparing SFB4 and SFB4', 4 base deletions at 903 bp were observed on the mutant. Subsequently, with a specially designed primer for SFB4', the sequencing indicated the SFB in 7 of the specimens to be SFB4'. Conclusion Six S genes were identified in the 39 C. avium specimens collected in Sichuan. The S4 from 7 of the specimens was the mutant gene, SFB4'. -
Key words:
- Sweet cherry /
- self-incompatibility /
- S-RNase /
- SFB
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图 1 部分甜樱桃材料基因组DNA凝胶电泳检测
Figure 1. Agarose gel electrophoresis of sweet cherry genomic DNA
图 2 通用引物对部分甜樱桃S-RNase 基因扩增结果
A～F分别为引物对BFP73/BFP74、BFP93/BFP94、Pru-C2/Pru-C4、Pru-C2/Pru-C3R、Pru-C2/Pru-C5R、PMT2/Pru-C3R扩增结果。
Figure 2. Agarose electrophoresis of S-RNase amplification of some sweet cherry specimens using universal primers
A–F: Agarose electrophoresis of primers, BFP73/BFP74, BFP93/BFP94, Pru-C2/Pru-C4R, Pru-C2/Pru-C3R, Pru-C2/Pru-C5R, and PMT2/Pru-C3R, respectively.
图 3 特异引物对部分甜樱桃S-RNase基因扩增将结果
A～F分别为引物PaS1F/PaS1R、PaS2F/PaS2R、PaS3F/PaS3R、PaS4F/PaS4R、PaS6F/PaS6R、PaS9F/PaS9R扩增结果。
Figure 3. Agarose electrophoresis of S-RNase amplification of some sweet cherry specimens using specific primers
A–F: Agarose electrophoresis of primers, PaS1F/PaS1R, PaS2F/PaS2R, PaS3F/PaS3R, PaS4F/PaS4R, PaS6F/PaS6R, and PaS9F/PaS9R, respectively.
图 5 SFB4'特异引物对11个含SFB4基因材料电泳图
Figure 5. Electropherogram of 11 SFB4-containing primer pairs with SFB4'-specific primers
表 1 39份甜樱桃材料信息
Table 1. information on 39 sweet cherry specimens
编号
Number名称
Name原产地
Origin编号
Number名称
Name原产地
Origin1 鲁樱3号 Luying 3 中国 21 龙冠 Longguan 中国 2 拉宾斯 Lapins 加拿大 22 佐藤锦 Satonishiki 日本 3 早甘阳 Zaoganyang 中国 23 贾红 Jiahong 中国 4 齐早 Qizao 中国 24 琥珀 Hupo 中国 5 鲁玉 Luyu 中国 25 桑德拉玫瑰 Sandra rose 加拿大 6 美早 Tieton 美国 26 彩玉 Caiyu 中国 7 布鲁克斯 Brooks 美国 27 罗亚明 Royal minnie 美国 8 桑提娜 Santina 加拿大 28 罗亚理 Royal lee 美国 9 萨米脱 Summit 加拿大 29 瑞德 Ruide 美国 10 黄蜜 Huangmi 中国 30 水晶香槟 Pearl champagne 美国 11 雷尼 Rainier 美国 31 珊瑚香槟 Coral champagne 美国 12 福星 Fuxing 中国 32 科迪亚 Kodia 捷克 13 明珠 Mingzhu 中国 33 那翁 Napoleon 德国 14 鲁樱1号 Luying 1 中国 34 艳阳 Sunburst 加拿大 15 黑珍珠 Heizhenzhu 中国 35 红蜜 Hongmi 中国 16 福晨 Fuchen 中国 36 大紫 Black tartarin 俄罗斯 17 早大果 Крупноплодная 乌克兰 37 早红宝石 Early ruby 乌克兰 18 俄罗斯8号 Russia 8 俄罗斯 38 宾库 Bing 美国 19 先锋 Van 加拿大 39 意大利早红 Italian early 法国 20 红灯 Hongdeng 中国 
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表 2 S基因引物名称及序列
Table 2. Name and sequence of S-RNase primer
名称
Name序列
Sequence名称
Name序列
SequencePru-C2 CTATGGCCAAGTAATTATTCAAACC PaS2R AAGTGCAATCGTTCATTTG Pru-C4R GGATGTGGTACGATTGAAGCG PaS3F GGGTCGCGATTTAAGAAAGAGC Pa-C5R CAAAATACCACTTCATGTAGCAACTG PaS3R AACAATCGTACTTTGTGATGACTTCG Pru-C3R TACCACTTCATGTAACAACTGAG PaS4F CACTGGGTCGCTGTTTAACTTTAGG PMT2 GCCTCTCCCATTCTGTTGTATTTC PaS4R TTGCATTTGATTAAGTGAGGCTTCA Pa-C3R TTGTATCATTGCCACTTTCCACG PaS6F ACTGGACCGCAATTTAAGCG BFP73 TGGCCAAGTAATTATTCAAACCC PaS6R AGTTGCTGCTTTAATGGGTGCA BFP74 CAAAATACCACTTCATGTAACAAC PaS9F TTTGTTACGTTATGAGCAGCAG BFP93 GTTCTTGCTTTTGCTTTCTTC PaS9R ATGAAACAATACATACCACTTIGCAT BFP94 CATAGGCCATGGATGGTG SFB4F ATGACATTCACACTACGTAAGAAAG PaS1F GTAATTGCAACGGGTCAAAATATGAG SFB4R TAAGTATTATTGAGTAAAACTAAAC PaS1R ACAACTCAGTATTAGTTGCTGGATCA PaS4'F TTAATGACTACAAGGCTGTAAGGA PaS2F CCTGCTTACTTTGTCACGCA PaS4'R CATAATAATGAGAAGGATAAAAGGAC
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表 3 39份甜樱桃材料S基因型
Table 3. S-RNase type of 39 sweet cherry specimens
编号
Number名称
NameS基因型
S-genotype编号
Number名称
NameS基因型
S-genotype1 鲁樱3号 Luying 3 S2S9 21 龙冠 Longguan S1S9 2 拉宾斯 Lapins S3S4 22 佐藤锦 Satonishiki S3S6 3 早甘阳 Zaoganyang S4S9 23 贾红 Jiahong S3S4 4 齐早 Qizao S4S9 24 琥珀 Hupo S1S2 5 鲁玉 Luyu S3S9 25 桑德拉玫瑰 Sandra rose S3S4 6 美早 Tieton S3S9 26 彩玉 Caiyu S1S3 7 布鲁克斯 Brooks S1S9 27 罗亚明 Royal minnie S1S3 8 桑提娜 Santina S1S4 28 罗亚理 Royal lee S3S6 9 萨米脱 Summit S1S2 29 瑞德 Ruide S1S3 10 黄蜜 Huangmi S3S6 30 水晶香槟 Pearl champagne S1S3 11 雷尼 Rainier S3S9 31 珊瑚香槟 Coral champagne S1S3 12 福星 Fuxing S1S3 32 科迪亚 Kodia S3S9 13 明珠 Mingzhu S6S9 33 那翁 Napoleon S3S4 14 鲁樱1号 Luying 1 S3S9 34 艳阳 Sunburst S3S4 15 黑珍珠 Heizhenzhu S1S4 35 红蜜 Hongmi S4S9 16 福晨 Fuchen S3S9 36 大紫 Black tartarin S6S9 17 早大果 Крупноплодная S3S9 37 早红宝石 Early ruby S3S9 18 俄罗斯8号 Russia 8 S3S6 38 宾库 Bing S3S6 19 先锋 Van S3S9 39 意大利早红 Italian early S3S4 20 红灯 Hongdeng S3S9
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