芝麻种间杂交亲和性差异及杂种生物学特征分析

doi:10.3864/j.issn.0578-1752.2022.20.003

中国农业科学 ›› 2022, Vol. 55 ›› Issue (20): 3897-3909.doi: 10.3864/j.issn.0578-1752.2022.20.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

芝麻种间杂交亲和性差异及杂种生物学特征分析

琚铭(),苗红梅,黄盈盈,马琴,王慧丽,王翠英,段迎辉,韩秀花,张海洋()

河南省农业科学院芝麻研究中心/河南省特色油料基因组学重点实验室/神农种业实验室,郑州 450002

收稿日期:2022-05-29 接受日期:2022-07-18 出版日期:2022-10-16 发布日期:2022-10-24
通讯作者: 张海洋
作者简介:琚铭,E-mail： jumingzz@163.com。
基金资助:
国家现代农业（特色油料）产业技术体系(CARS-14);河南省重大科技专项(201300110600);中原科学家工作室建设项目(092101211100);中原科学家工作室建设项目(214400510026);中原科技创新领军人才项目(214200510020);河南省农业科学院优秀青年项目(2020YQ19);科技创新团队项目(2022TD05);科技创新创意项目(2020CX02);自主创新项目(2021ZC70)

Analysis of Cross Compatibility Variation Among Diverse Sesamum Species and Biological Characteristics of the Interspecific Hybrid Progenies

JU Ming(),MIAO HongMei,HUANG YingYing,MA Qin,WANG HuiLi,WANG CuiYing,DUAN YingHui,HAN XiuHua,ZHANG HaiYang()

Henan Sesame Research Center, Henan Academy of Agricultural Sciences/Henan Key Laboratory of Specific Oilseed Crops Genomics/Shennong Laboratory, Zhengzhou 450002

Received:2022-05-29 Accepted:2022-07-18 Online:2022-10-16 Published:2022-10-24
Contact: HaiYang ZHANG

摘要/Abstract

摘要：

【目的】探索芝麻不同种之间的杂交亲和性,分析其杂种的生物学特征,为芝麻野生种种质资源高效利用提供依据。【方法】以芝麻栽培种豫芝11号（S. indicum,2n=26）和S. latifolium（2n=32）、S. calycinum（2n=32）、S. angustifolium（2n=32）、S. radiatum（2n=64）等4个野生种为亲本材料,采用双列杂交方法,通过田间人工授粉配置不同种间组合;结合胚拯救方法获得种间杂种F₁。根据杂交结蒴率比较组合杂交亲和性;在盛花期和成熟期观察杂种植物学性状特征,利用Alexander染色法进行花粉粒育性鉴定。通过根尖细胞染色体涂片明确杂种染色体数目及特征。选用自主筛选的胡麻属特异多态性SSR引物,分析种间杂种分子标记差异。【结果】配置了5个芝麻种间的20个正、反交组合,共授粉2 091朵花,获得杂交蒴果370个。发现以染色体数目多的种为母本更易获得远缘杂交蒴果。5个芝麻种之间杂交亲和性的变化范围为1.18%（S. radiatum×S. calycinum）—63.33%（S. calycinum×S. angustifolium）。共有9个杂交组合获得杂种F₁种子,F₁植株的花粉败育率为35.21%—100.00%,其中,S. calycinum与S. angustifolium杂交组合F₁的可育株比例最高,为87.68%。杂种F₁在株高、株型等性状方面均表现出明显的超亲优势。栽培种与各野生种的正反交杂种F₁在叶型、花型和花色表现出双亲的局部特征。栽培种芝麻（n=13）与具有n=16染色体组型的3个野生种的杂交亲和性依次为S. angustifolium>S. calycinum>S. latifolium;野生种S. radiatum（n=32）与n=16染色体组的3个野生种的亲和性依次为S. calycinum>S. angustifolium>S. latifolium。在5个种中,野生种S. calycinum与S. angustifolium的亲缘关系相对最近。获得的部分杂种植株根尖细胞染色体数目观察显示,杂种的染色体数目与理论值一致。利用3对多态性SSR引物对F₁植株的分子鉴定结果显示,真杂种比例为99.66%。杂种染色体核型和特异SSR标记结果显示出胡麻属不同种的遗传特征差异。【结论】胡麻属5个种之间的杂交亲和性差异显著,种间杂交后代杂种优势明显;S. calycinum与S. angustifolium的亲缘关系相对最近,可用于芝麻优异种质创制和远缘杂交育种研究;其他种间杂交存在着生殖隔离障碍,可采用胚拯救、分子标记利用等手段加强芝麻野生资源利用。

关键词: 芝麻, 野生种, 种间杂交, 生物学特征, 分子鉴定

Abstract:

【Objective】 The research aims to explore the cross compatibility between different Sesamum species and analyze the biological characteristics of interspecies hybrid progeny so as to supply the foundation for efficient application of wild sesame species. 【Method】 A sesame cultivar Yuzhi 11 (S. indicum, 2n=26) and 4 wild Sesamum species including S. latifolium (2n=32), S. calycinum (2n=32), S. angustifolium (2n=32), and S. radiatum (2n=64) were applied to construct interspecies cross combinations using diallel hybridization method by artificial pollination in the field. Embryo rescue method was also used to obtain interspecific hybrid F₁. Interspecific hybrid compatibility was compared based on hybrid capsule formation rate. Botanical characters of hybrids were observed during flowering and mature stages. Pollen fertility was assessed using Alexander staining method. Chromosome number and karyotype characteristics of root somatic cells of hybrids were observed using smear chromosome preparation technique. Specific and polymorphic SSR primers in Sesamum were used to analyze the molecular difference in interspecific hybrids.【Result】 Twenty positive and reciprocal cross combinations were constructed for the 5 Sesamum species. A total of 2091 flowers were pollinated and 370 hybrid capsules were harvested. As to the female parents with more chromosomes, hybrid capsules were more easily obtained. The cross compatibility among the 5 Sesamum species significantly varied from 1.18% (S. radiatum×S. calycinum) to 63.33% (S. calycinum×S. angustifolium). F₁ plants of 9 combinations produced hybrid seeds, while the ratio of pollen sterility of F₁ progeny ranged from 35.21%-100.00%. The cross S. calycinum×S. angustifolium presented the highest sterility ratio to 87.68%. Hybrid progeny exhibited the obvious heterosis over parents in plant height, plant type, and some key agronomic traits. As to the positive and reciprocal hybrid F₁ derived from sesame cultivar and the wild species, leaf shape, flower shape, and flower color showed partial characters of both parents. The cross compatibility between sesame cultivar (n=13) and the 3 Sesamum species with chromosome group n=16 ranked as S. angustifolium>S. calycinum>S. latifolium. The cross compatibility between wild species S. radiatum (n=32) and the 3 species with n=16 ranked as S. calycinum>S. angustifolium>S. latifolium. Among the 5 Sesamum species, the genetic relationship between S. calycinum and S. angustifolium is relatively closest. The chromosome number of root tip cells of some hybrid plants is consistent with the theoretical value calculated from the parents. Screening results of the 3 pairs of polymorphic SSR primers indicated that 99.66% of obtained F₁ plants are true hybrid. Chromosome karyotype and SSR marker screening results reflected the genetic difference and characters of Sesamum species. 【Conclusion】Among the 5 Sesamum species, the cross compatibility varies significantly and the heterosis of interspecific hybrid is obvious. Of which only S. calycinum and S. angustifolium have the relatively closest genetic relationship and could be directly applied for elite germplasm creation and interspecific hybrid breeding in Sesamum. Reproductive isolation barriers exist in other cross combinations. Some techniques including embryo rescue and molecular marker application should be used to achieve the utilization of wild Sesamum species for sesame breeding.

Key words: sesame, wild Sesamum species, interspecific hybridization, botanic character, molecular assessment

琚铭, 苗红梅, 黄盈盈, 马琴, 王慧丽, 王翠英, 段迎辉, 韩秀花, 张海洋. 芝麻种间杂交亲和性差异及杂种生物学特征分析[J]. 中国农业科学, 2022, 55(20): 3897-3909.

JU Ming, MIAO HongMei, HUANG YingYing, MA Qin, WANG HuiLi, WANG CuiYing, DUAN YingHui, HAN XiuHua, ZHANG HaiYang. Analysis of Cross Compatibility Variation Among Diverse Sesamum Species and Biological Characteristics of the Interspecific Hybrid Progenies[J]. Scientia Agricultura Sinica, 2022, 55(20): 3897-3909.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2022.20.003

https://www.chinaagrisci.com/CN/Y2022/V55/I20/3897

图/表 10

表1

表2

表3

种间杂交组合结实性和杂种胚拯救出苗率统计"

编号 No.	种间杂交组合 Interspecific hybrid combination	母本 Female (♀)	父本 Male (♂)	杂交结实性 Hybrid capsule formation						胚拯救 Hybrid embryo rescue
编号 No.	种间杂交组合 Interspecific hybrid combination	母本 Female (♀)	父本 Male (♂)	授粉花朵数 Pollinated flower <BOLD>N</BOLD>o.	结蒴数量 Capsule <BOLD>N</BOLD>o.	杂交结蒴率 Capsule ratio (%)	种子数量 Hybrid seed <BOLD>N</BOLD>o.	杂种出苗数 Hybrid seedlings <BOLD>N</BOLD>o.	杂种出苗率 Hybrid plantlet ratio (%)	幼胚数量 Young embryo <BOLD>N</BOLD>o.	组培苗数量 Seedling <BOLD>N</BOLD>o.	成苗率 Seedling ratio (%)
H1a	S. indicum×S. latifolium	豫芝11号 Yuzhi 11	Ken1	60	9	15.00	56	0	0	105	24	22.86
H1b	S. latifolium×S. indicum	Ken1	豫芝11号 Yuzhi 11	70	11	15.71	255	0	0	266	186	69.92
H2a	S. indicum×S. calycinum	豫芝11号 Yuzhi 11	Ken8	110	20	18.18	150	0	0	156	2	1.28
H2b	S. calycinum×S. indicum	Ken8	豫芝11号 Yuzhi 11	90	3	3.33	47	1	2.13	375	87	23.00
H3a	S. indicum×S. angustifolium	豫芝11号 Yuzhi 11	G01	40	9	22.50	156	0	0	75	1	1.30
H3b	S. angustifolium×S. indicum	G01	豫芝11号 Yuzhi 11	320	15	4.69	210	1	0.48	112	5	4.46
H4a	S. indicum×S. radiatum	豫芝11号 Yuzhi 11	G02	249	82	32.93	57	0	0	225	16	7.11
H4b	S. radiatum×S. indicum	G02	豫芝11号 Yuzhi 11	253	104	41.11	78	2	2.56	350	51	14.57
H5a	S. latifolium×S. calycinum	Ken1	Ken8	30	3	10.00	78	0	0	130	0	0
H5b	S. calycinum×S. latifolium	Ken8	Ken1	102	8	7.84	137	0	0	85	3	3.52
H6a	S. angustifolium×S. latifolium	G01	Ken1	45	5	11.11	50	0	0	75	8	11.11
H6b	S. latifolium×S. angustifolium	Ken1	G01	51	15	29.41	236	3	1.27	50	1	2.00
H7a	S. latifolium×S. radiatum	Ken1	G02	57	6	10.53	55	0	0	440	5	1.14
H7b	S. radiatum×S. latifolium	G02	Ken1	64	10	15.63	200	0	0	300	2	0.60
H8a	S.calycinum×S. angustifolium	Ken8	G01	30	19	63.33	285	44	15.44	105	98	93.00
H8b	S.angustifolium×S. calycinum	G01	Ken8	25	12	48.00	105	26	24.75	150	66	44.00
H9a	S. calycinum×S. radiatum	Ken8	G02	56	2	3.57	12	0	0	125	0	0
H9b	S. radiatum×S. calycinum	G02	Ken8	255	3	1.18	20	0	0	120	33	27.50
H10a	S. angustifolium×S. radiatum	G01	G02	99	5	5.05	55	0	0	0	0	0
H10b	S. radiatum×S. angustifolium	G02	G01	85	29	34.12	909	0	0	280	22	7.86

表3

图1

图2

图3

表4

芝麻不同种及种间杂交F1植株的生物学特征统计"

材料名称 Name	来源 Source	叶型 Leaf type	株高 Plant height (cm)	株型 Plant type	蜜腺 Honey gland	腋叶花数 flower no. per axillar	花长 Corolla length (cm)	花宽 Corolla width (cm)	花冠颜色 Corolla color	花舌颜色 Corolla tip color	花粉败育率 Pollen grain fertility (%)	自交结蒴性 Self- pollination capsule formation	有无成熟种胚 Embryo in mature seed	种皮颜色 Seed color
豫芝11号 Yuzhi 11	S. indicum	椭圆形 Oval	169.2	直立、单秆 Erect, uniculm	无 Without	3	3.7	2.4	白色 White	白色 White	0	是 Yes	有 Have	白色 White
Ken1	S. latifolium	心形 Heart-shaped	230.5	直立、强分支 Erect, Many branches	黄色 Yellow	1	4.5	2.4	淡紫色 Lavender	紫条 Purple strip	2.22	是 Yes	有 Have	黑色 Black
Ken8	S. calycinum	条形 Strip type	151.6	半匍匐、强分支 Semi-erect, many branches	黄色 Yellow	1	3.9	2.0	紫色 Purple	淡紫色 Lavender	0	是 Yes	有 Have	黑色 Black
G01	S. angustifolium	条形 Strip type	122.6	直立、弱分支 Erect, few branches	黄色 Yellow	1	3.7	2.0	紫红色 Mauve	紫红色 Mauve	1.05	是 Yes	有 Have	黑色 Black
G02	S. radiatum	披针形 lanceolate	179.6	直立、弱分支 Erect, few branches	紫色 Purple	1	4.2	2.4	白色 White	白色 White	4.28	是 Yes	有 Have	黑色 Black
H1a	S. indicum× S. latifolium	条形 Strip type	240.2	直立、强分支 Erect, strong branch	黄色 Yellow	1	5.0	3.0	淡紫色 Lavender	粉色条纹 Pink stripes	100.00	是 Yes	无 No	黑色 Black
H1b	S. latifolium× S. indicum	条形 Strip type	251.7	直立、强分支 Erect, strong branch	黄色 Yellow	1	4.5	2.5	淡紫色 Lavender	紫色条纹 Purple stripes	100.00	是 Yes	无 No	黑色 Black
H2b	S. calycinum× S. indicum	条形 Strip type	171.5	半匍匐、强分支 Semi-erect, many branches	黄色 Yellow	1	4.0	2.6	紫色 Purple	紫色条纹 Purple stripes	97.78	否 No	/	/
H4b	S. radiatum× S. indicum	披针形 lanceolate	186.9	直立、强分支 Erect, strong branch	紫色 Purple	1	4.1	2.2	粉色 Pink	白色 White	100.00	是 Yes	无 No	黑色 Black
H5a	S. latifolium× S. angustifolium	卵圆形 Ovate	191.4	直立、弱分支 Erect, few branches	黄色 Yellow	1	4.2	2.1	粉色 Pink	粉色有紫色纹路 Pink with purple texture	100.00	否 No	/	/
H7a	S. calycinum× S. angustifolium	条形 Strip type	170.1	半匍匐、强分支 Semi-erect, many branches	紫色 Purple	1	4.0	2.5	紫色 Purple	紫色 Purple	68.89	否 No	有 Have	黑色 Black
H7b	S. angustifolium× S. calycinum	条形 Strip type	169.4	半匍匐、强分支 Semi-erect, many branches	紫色 Purple	1	4.0	2.5	紫色 Purple	紫色 Purple	35.21	否 No	有 Have	黑色 Black
H8b	S. radiatum× S. calycinum	披针形 Lanceolate	199.8	半匍匐、强分支 Semi-erect, many branches	紫色 Purple	1	4.5	2.5	紫色 Purple	紫色条纹 Purple stripes	100.00	否 No	/	/
H9b	S. radiatum× S. angustifolium	柳叶型 Willow leaf shape	200.3	直立、弱分支 Erect, few branches	深紫色 Modena	1	4.0	2.2	紫色 Purple	白色有紫色边缘 White with purple edges	100.00	否 No	/	/

表4

图4

图5

表5

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引物Primer	引物序列Primer sequences (5′-3′)
HS94	F：CATGTGTTCTCTCCCACCAC
HS94	R：TCTTGACCATGTTTTCCACC
HS352	F：TTCCGCTGCTTGTATGATTC
HS352	R：TGGTGGAAAAAGAAGGGAAC
HS53	F：GAAGCTTGAAGAGAGGAGGG
HS53	R：ATGGAACTTCTCCGATCACC

芝麻种间杂交亲和性差异及杂种生物学特征分析

Analysis of Cross Compatibility Variation Among Diverse Sesamum Species and Biological Characteristics of the Interspecific Hybrid Progenies

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材料名称 Material name	种属特性 Sesamum species	体细胞染色体数目 Chromosome number in somatic cell	株型 Plant type
豫芝11号Yuzhi 11	S. indicum	2n=26	直立、单秆 Erect, uniculm
Ken1	S. latifolium	2n=32	直立、分支 Erect, branching
Ken8	S. calycinum	2n=32	半匍匐、分支 Semi-erect, branching
G01	S. angustifolium	2n=32	直立、分支 Erect, branching
G02	S. radiatum	2n=64	直立、分支 Erect, branching

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