东试早柚与沙田柚和水晶柚遗传背景比较

doi:10.16420/j.issn.0513-353x.2022-0981

园艺学报 ›› 2023, Vol. 50 ›› Issue (11): 2337-2349.doi: 10.16420/j.issn.0513-353x.2022-0981

• 遗传育种·种质资源·分子生物学 • 上一篇下一篇

东试早柚与沙田柚和水晶柚遗传背景比较

刘承浪¹^,^*, 冯迪¹^,^*, 曹宗洪¹, 闫素云³, 张永青¹, 徐祥增², 高世德², 叶俊丽¹, 柴利军¹, 谢宗周¹^,^**(), 邓秀新¹^,^**()

¹ 华中农业大学园艺林学学院，园艺植物生物学教育部重点实验室，武汉 430070
² 云南省热带作物科学研究所，云南景洪 666100
³ 云南省农业科学院热带亚热带经济作物研究所，云南保山 678000

收稿日期:2023-05-31 修回日期:2023-08-28 出版日期:2023-11-25 发布日期:2023-11-28
通讯作者:
**（E-mail：xiezz@webmail.hzau.edu.cn，
xxdeng@mail.hzau.edu.cn）
作者简介:
*共同第一作者
基金资助:
云南省科技厅重大科技专项(202102AE090054); 国家重点研发计划项目(2019YFD1001400)

Comparative Analysis of Genetic Background of‘Dongshi Zaoyou’，‘Shatianyou’and‘Shuijingyou’Pummelos

LIU Chenglang¹^,^*, FENG Di¹^,^*, CAO Zonghong¹, YAN Suyun³, ZHANG Yongqing¹, XU Xiangzeng², GAO Shide², YE Junli¹, CHAI Lijun¹, XIE Zongzhou¹^,^**(), DENG Xiuxin¹^,^**()

¹ Key Laboratory of Horticultural Plant Biology（Ministry of Education），College of Horticulture and Forestry Sciences，Huazhong Agricultural University，Wuhan 430070，China
² Yunnan Institute of Tropical Crops，Jinghong，Yunnan 666100，China
³ Institute of Tropcal and Subtropical Cash Crops，Yunnan Academy of Agricultural Scienles，Baoshan，Yunnan 678000，China

Received:2023-05-31 Revised:2023-08-28 Published:2023-11-25 Online:2023-11-28
Contact:
**（E-mail：xiezz@webmail.hzau.edu.cn，
xxdeng@mail.hzau.edu.cn）

摘要/Abstract

摘要：

‘东试早柚’是实生选育出的柚品种，在云南省热带地区种植，具有早熟、丰产、无籽等优点，然而其来源至今尚未厘清，在品种权意识淡薄的栽培地区产生了许多经济纠纷。因此，解析‘东试早柚’的遗传来源对保护和利用该品种均具有重要意义。以西双版纳‘东试早柚’及其相近品种‘沙田柚’‘水晶柚’为材料，利用分子标记和生物信息等手段探究了3个品种的遗传背景差异。结果显示，‘东试早柚’与‘水晶柚’间杂交不亲和，而与‘沙田柚’杂交亲和；‘东试早柚’和‘水晶柚’的自交不亲和S位点基因型均为S₅S₆，而‘沙田柚’为S₁S₂；‘东试早柚’和‘水晶柚’全基因组SNP密度分布高度相似，SNP聚类分析将‘东试早柚’和‘水晶柚’聚为一类，而与‘沙田柚’显著区分。此外，2对InDel多态性标记也可明显区分‘东试早柚’与‘沙田柚’。因此，本研究结果认为‘东试早柚’与‘水晶柚’遗传背景更为相近，而与‘沙田柚’遗传距离较远、相差很大。

关键词: 东试早柚, S基因型, InDel标记, SNP

Abstract:

‘Dongshi Zaoyou’，a new pummelo cultivar selected from pummelo seedlings，is mainly cultivated in the tropical areas of Yunnan Province. It has good characteristics such as early-maturing，high-yielding，seedless and etc. However，the genetic origin of‘Dongshi Zaoyou’pummelo has not yet been clarified，which was unfavorable to its protection and utilization. In this study，the cross-compatibility test and molecular markers including self-incompatibility S-genotype，SNP and InDels were used to explore the genetic background differences between‘Dongshi Zaoyou’pummelo and its two similar cultivars ‘Shatianyou’，‘Shuijingyou’. The pollination experiments revealed the cross-incompatibility between‘Dongshi Zaoyou’pummelo and‘Shuijingyou’pummelo while the cross-compatibility between‘Dongshi Zaoyou’pummelo and‘Shatianyou’pummelo. Accordingly，S-genotype of‘Dongshi Zaoyou’pummelo was judged to be S₅S₆，the same as that of‘Shuijingyou’pummelo（S₅S₆），while different from that of ‘Shatianyou’pummelo（S₁S₂）. Based on the genome resequencing data，the genome-wide SNP density distribution of three pummelo cultivars were then plotted and compared. The results showed that‘Dongshi Zaoyou’pummelo and‘Shuijingyou’pummelo had highly similar SNP density and clustered into a group，but they both were significantly different from‘Shatianyou’pummelo. In addition，two InDel polymorphic markers could clearly distinguish‘Dongshi Zaoyou’pummelo from‘Shatianyou’pummelo. Collectively，it could be concluded that‘Dongshi Zaoyou’pummelo showed a closer relationship to‘Shuijingyou’pummelo than‘Shatianyou’pummelo.

Key words: ‘Dongshi Zaoyou’pummelo, S-genotype, InDel marker, SNP

刘承浪, 冯迪, 曹宗洪, 闫素云, 张永青, 徐祥增, 高世德, 叶俊丽, 柴利军, 谢宗周, 邓秀新. 东试早柚与沙田柚和水晶柚遗传背景比较[J]. 园艺学报, 2023, 50(11): 2337-2349.

LIU Chenglang, FENG Di, CAO Zonghong, YAN Suyun, ZHANG Yongqing, XU Xiangzeng, GAO Shide, YE Junli, CHAI Lijun, XIE Zongzhou, DENG Xiuxin. Comparative Analysis of Genetic Background of‘Dongshi Zaoyou’，‘Shatianyou’and‘Shuijingyou’Pummelos[J]. Acta Horticulturae Sinica, 2023, 50(11): 2337-2349.

图/表 11

表1 ‘东试早柚’遗传鉴定所用的InDel引物信息

Table1 InDel primers used in the genetic identification of‘Dongshi Zaoyou’pummelo

引物名称 Primer name	引物序列（5′-3′） Sequence	退火温度/℃ Annealing temperature
CH1	F：ATGAATGCTTGTGCAAGGACAGCTAAATTC；R：TTAAAGATTTTCCATCTGGGTAAAAGCAT	56
CH2	F：GAGGGCAGCAGTTTGAATCC；R：TGGGACCAACTTACATCAATGC	55
CH3	F：CGTTCGAGTCGTCTTTGTTGG；R：ATACACTGTAAGAGAGATCGTAACA	56
CH4	F：ATGAATGCTTGTGCAAGGACAGCTAAATTCCAG；R：TTAAAGATTTTCCATCTGGGTAAAAGCATCTTC	55
CH5	F：CGCGGAGTCAGCAACAAAAA；R：ACACTGTAAGAGAGATCGTAACAT	55
CH6	F：AGCAACCCGAAACACAAACAC；R：ACACTGTAAGAGAGATCGTAACATT	55
CH7	F：GGATTTGGCTTTTCATCTTCTCA；R：TGCTAATGAGAGATAGTGGGGAG	56

表2 S基因型鉴定的特异性引物

Table 2 The specific primers for S-genotypes identification

引物名称 Primer name	引物序列（5′-3′） Sequence	退火温度/℃ Annealing temperature
S₁	F：ATGAACATTACTTTCTTCCTCTA；R：TTAAGGCGGGGGAAAGGTA	55
S₂	F：ATGATATCGACAAAGACGAAAA；R：CTACTCATCGGTCGGCTCG	55
S₅	F：ATGAAGGTGGCATCCATCAAC；R：CTACCACCGTGGTGGGAAAATAATATCC	56
S₆	F：ATGGGGACTAATTTCCTCATTATC；R：CTATATTTTAACGTATCGCGGC	55

图1 不同柚品种的表型特征

Fig. 1 Phenotypic characteristics of different pommelo cultivars

表3 不同柚品种果实品质

Table 3 Fruit quality of different pummelos

试验材料 Plant material	单果质量/g Fruit weight	纵径/mm Vertical diameter	横径/mm Transverse diameter	果形指数 Fruit shape index
‘东试早柚’Dongshi Zaoyou	1 062.17 ± 25.34 bc	151.18 ± 9.01 b	143.66 ± 2.55 b	1.05 ± 0.06 ab
‘沙田柚’Shatianyou	1 543.60 ± 165.96 a	192.06 ± 8.96 a	160.61 ± 10.74 a	1.20 ± 0.05 a
‘水晶柚1’Shuijingyou 1	1 207.26 ± 63.46 bc	152.99 ± 1.35 b	144.49 ± 6.25 b	1.06 ± 0.05 ab
‘水晶柚2’Shuijingyou 2	1 048.00 ± 123.57 c	155.99 ± 14.24 b	149.95 ± 4.19 ab	1.04 ± 0.09 b
‘水晶柚3’Shuijingyou 3	1 349.62 ± 204.80 ab	187.94 ± 10.11 a	158.14 ± 7.81 ab	1.19 ± 0.06 a
试验材料 Plant material	可溶性固形物/% Total soluble solid	可滴定酸/% Titratable acid	固酸比 Solidity-acid ratio	维生素C/（mg · kg^-1） Vitamin C
‘东试早柚’Dongshi Zaoyou	10.93 ± 0.19 a	0.16 ± 0.02 b	71.36 ± 9.49 a	788.8 ± 1.32 a
‘沙田柚’Shatianyou	8.55 ± 0.70 b	0.41 ± 0.18 a	27.20 ± 15.82 b	646.0 ± 8.47 b
‘水晶柚1’Shuijingyou 1	9.44 ± 0.40 b	0.32 ± 0.07 ab	30.32 ± 5.44 b	570.7 ± 4.57 b
‘水晶柚2’Shuijingyou 2	8.95 ± 0.24 b	0.18 ± 0.03 ab	52.51 ± 10.48 b	568.2 ± 2.71 b
‘水晶柚3’Shuijingyou 3	8.88 ± 0.62 b	0.25 ± 0.05 ab	37.00 ± 5.35 b	539.6 ± 6.22 b

图2 不同柚品种杂交授粉后花柱的荧光成像

Fig. 2 Fluorescence images of pollen tubes in pistils after different pollinations

图3 不同柚品种S基因型鉴定

Fig. 3 S-genotypes of different pummelo cultivars S1，S2，S5，S6：S1-RNase，S2-RNase，S5-RNase，S6-RNase.

图4 ‘东试早柚’（DS）与’水晶柚’（SJ）S5-RNase（A）、S6-RNase（B）序列比对

Fig. 4 Sequence alignment of S5-RNase（A）and S6-RNase（B）between‘Dongshi Zaoyou’（DS）and‘Shuijingyou’（SJ）pummelo

图5 不同柚SNP密度对比 A、B、C：分别是‘东试早柚’‘水晶柚’‘沙田柚’SNP密度直方图；D：染色体；E、F、G：分别是‘东试早柚’‘水晶柚’‘沙田柚’SNP密度热图。

Fig. 5 Comparison of SNP density in different pummelo cultivars A，B and C：SNP density histograms of‘Dongshi Zaoyou’，‘Shuijingyou’and‘Shatianyou’pummelo，respectively；D：Chromosome；E，F and G：SNP density heat maps of‘Dongshi Zaoyou’，‘Shuijingyou’and‘Shatianyou’pummelo，respectively.

图6 SNP变异位点数差值

Fig. 6 Number difference of SNP variant loci

图7 ‘东试早柚’、‘沙田柚’和‘水晶柚’的进化树图中数值表示枝长信息，即遗传变异度；Tree scale表示遗传变异度为0.1指示的枝长。

Fig. 7 The evolutionary trees of‘Dongshi Zaoyou’，‘Shatianyou’and‘Shuijingyou’pummelo The values in the graph indicate the branch length information，i.e. genetic variability；Tree scale denotes branch lengths with a genetic variance of 0.1 indication.

图8 InDel标记（CH1 ~ CH7）在‘东试早柚’（DS）与‘沙田柚’（ST）中的扩增

Fig. 8 Amplification of InDel markers in‘Dongshi Zaoyou’（DS）and‘Shatianyou’（ST）pummelo

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东试早柚与沙田柚和水晶柚遗传背景比较

Comparative Analysis of Genetic Background of‘Dongshi Zaoyou’，‘Shatianyou’and‘Shuijingyou’Pummelos

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东试早柚与沙田柚和水晶柚遗传背景比较

Comparative Analysis of Genetic Background of‘Dongshi Zaoyou’，‘Shatianyou’and‘Shuijingyou’Pummelos

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