利用胚挽救技术创制无核抗寒葡萄新种质

doi:10.3864/j.issn.0578-1752.2018.21.010

Abstract

Abstract:

【Objective】 The objective of this study is to obtain new cold-resistant seedless grape germplasm and improve the breeding efficiency through embryo rescue in vitro. 【Method】 The different genotypes, basal culture media (MM3 and ER), amino acids (2.5 mmol·L ^-1cysteine, asparagine, glutamine and serine were added in MM3 medium) were used to investigate the effect on the embryo development rates and seedling rates from ovules of 7 cross combinations. From the parents, the molecular markers GSLP1-569 and SCC8-1018 for seedlessness and the marker S241-717 for cold-resistance were used to identify the strains with specific bands, and then the three markers were used to screen the hybrids by recognizing the specific bands. 【Result】 Of the 7 combinations, 1 168 ovules cultured in MM3 were obtained 331 embryos and 97 hybrid strains. Among these crosses, ‘Ruby Seedless’ × ‘Beichun’ got the highest embryo development rate and seedling rate, which were 46.00% and 17.33%, respectively. Compared with female parents, for ‘Shuangyou’ as male parent and ‘Blush Seedless’ ‘Thompson Seedless’ ‘Kunxiang Seedless’ as female parents, ‘Kunxiang Seedless’ × ‘Shuangyou’ had the highest embryo development rate and seedling rate, which were 45.39% and 16.31%, respectively. For ‘Beibinghong’ as male parent and ‘Blush Seedless’ ‘Beauty Seedless’ ‘Thompson Seedless’ as female parents, ‘Blush Seedless’ × ‘Beibinghong’ had the highest embryo development rate and seedling rate, which were 33.52% and 6.59%, respectively. Different male parents also influenced seedlings formation. The embryo development rates of ‘Blush Seedless’ × ‘Beibinghong’ and ‘Blush Seedless’ × ‘Shuangyou’ were 33.52% and 39.00%, and the seedling rates of them were 6.59% and 9.50%, respectively. The embryo development rates of ‘Thompson Seedless’ × ‘Beibinghong’ and ‘Thompson Seedless’ × ‘Shuangyou’ were 7.64% and 19.26%, and the seedling rates of them were 1.91% and 8.15%, respectively. The rate of embryo development and seedling of ovule cultured in MM3 medium was higher than that of ER medium. For ‘Kunxiang Seedless’ × ‘Shuangyou’, the ovules cultured in MM3 medium with 2.5 mmol·L ^-1 glutamine showed the highest embryo development rate, which was 59.69%. The seedling rate of ovules cultured in basal media with asparagine and glutamine was 21.43% and 20.93%, respectively. The effect of promoting seedlings development was significant. For ‘Ruby Seedless’ × ‘Beichun’, the ovules in media with 2.5 mmol·L ^-1 asparagine showed the highest embryo development rate, which was 55.71%. The seedling rate of ovules cultured in basal media with asparagine and glutamine was 21.43% and 22.22%, respectively. The effect of promoting seedlings development was significant. By using the molecular markers GSLP1-569 and SCC8-1018 for seedlessness and the marker S241-717 for cold-resistance, 83 hybrid strains from 6 cross combinations were detected, and there were 49 strains with specific bands of the molecular markers linked to the seedlessness, 55 strains with specific bands of the markers S241-717 for cold-resistance. Importantly, 36 strains with both seedlessness and cold-resistance were identified in this study. 【Conclusion】 ‘Kunxiang Seedless’ and ‘Blush Seedless’ are suitable as the female parents for embryo rescue in vitro of seedless grapes. The embryo rescue efficiency of ‘Shuangyou’ as the male parent is higher than that of ‘Beibinghong’. MM3 is suitable for embryo rescue in vitro, and the addition of amide acids to MM3 culture medium is helpful to the seedling formation. Among the 83 hybrid strains detected by molecular markers, 36 strains may have the seedless and cold-resistant characteristics.

Key words: seedless grape, cold-resistant, embryo-rescue, ovule culture, molecular marker assisted selection

YaNan ZHAO,QiangWei LUO,YueJin WANG. Breeding for Grape Germplasm Involved in Seedlessness with Cold-Resistant Using Embryo Rescue[J].Scientia Agricultura Sinica, 2018, 51(21): 4119-4130.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2018.21.010

https://www.chinaagrisci.com/EN/Y2018/V51/I21/4119

Figures/Tables 11

Table 1

Table 2

Table 3

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Fig. 7

Fig. 8

References 33

[1]	唐冬梅 . 无核葡萄杂交胚挽救新种质创建与技术完善[D]. 杨凌: 西北农林科技大学, 2010.
	TANG D M . Novel germplasm innovation of seedless grapes by embryo rescue and technique improvement[D]. Yangling: Northwest A&F University, 2010. ( in Chinese)
[2]	彭冬燕, 袁媛, 李丹桐, 杨燕, 陈升东 . 世界水果生产概况. 西南农业学报, 2006. 19(增刊):505-507.
	PENG D Y, YUAN Y, LI D T, YANG Y, CHEN S D . Introduction to global fruit’s production. Southwest China Journal of Agricultural Sciences, 2006,19(Suppl.):505-507. (in Chinese)
[3]	王跃进, 万怡震 . 美国加州的葡萄生产与科研. 西北农林科技大学学报 (自然科学版), 2002,30(1):134-140. doi: 10.3321/j.issn:1671-9387.2002.01.032
	WANG Y J, WAN Y Z . Review of grape production and researches in California of US. Journal of Northwest A&F University (Nature Science Edition), 2002,30(1):134-140. (in Chinese) doi: 10.3321/j.issn:1671-9387.2002.01.032
[4]	潘学军 . 无核抗病葡萄胚挽救技术体系优化及新品系培育[D]. 杨凌: 西北农林科技大学, 2005.
	PAN X J . Innovating in the technique system of embryo rescue of stenospermocarpic grape and breeding new cultivars of both seedless and disease-resistance traits[D]. Yangling: Northwest A&F University, 2005. ( in Chinese)
[5]	郭海江 . 抗病无核葡萄新种质创制及分子标记辅助育种[D]. 杨凌: 西北农林科技大学, 2005.
	GUO H J . Germplasm innovation of disease resistant and seedless grape and molecular maker-assisted breeding[D]. Yangling: Northwest A&F University, 2005. ( in Chinese)
[6]	刘巧 . 利用胚挽救技术培育抗寒无核葡萄新种质[D]. 杨凌: 西北农林科技大学, 2015.
	LIU Q . Breeding new, cold-resistant, seedless grape germplasm using embryo rescue technology[D]. Yangling: Northwest A&F University, 2015. ( in Chinese)
[7]	徐丽莹, 张怀明, 金明秋 . 100PF-A型葡萄越冬覆土机性能及效益分析. 农业机械, 2004(4):52.
	XU L Y, ZHANG H M, JIN M Q . Performance and efficiency analysis of 100PF-A grape overwintering machine. Farm Machinery, 2004(4):52. (in Chinese)
[8]	RAMMING D W, EMERSHAD R L . In ovulo embryo culture of seeded and seedless Vitis vinifera(Abstr). HortScince, 1982,17(3):487.
[9]	EMERSHAD R L, RAMMING D W, SERPE M D . In ovulo embryo development and plant formation from stenospermic genotypes of Vitis vinifera. American Journal of Botany, 1989,76(3):397-402.
[10]	EMERSHAD R L, RAMMING D W . Somatic embryogenesis and plant development from immature zygotic embryos of seedless grapes ( Vitis vinifera L.). Plant Cell Reports, 1994,14(1):6-12. doi: 10.1007/BF00233289 pmid: 24194218
[11]	唐冬梅, 王跃进, 赵荣华, 潘学军, 蔡军社, 张剑侠, 张朝红, 骆强伟 . 无核葡萄胚挽救中影响胚发育的因子. 中国农业科学, 2009,42(7):2449-2457.
	TANG D M, WANG Y J, ZHAO R H, PAN X J, CAI J S, ZHANG J X, ZHANG C H, LUO Q W . Factors influencing embryo development in embryo rescue of seedless grapes. Scientia Agricultura Sinica, 2009,42(7):2449-2457. (in Chinese)
[12]	CAIN D W, ENERSHAD R L, TARAILO R E . In-ovulo embryo culture and seedling development of seeded and seedless grapes(Vitis vinifera L.). Vitis, 1983,22:9-14.
[13]	EMERSHAD R L, RAMMING D W . In-ovulo embryo culture of Vitis vinifera L.c.v. ‘Thompson Seedless’. American Journal of Botany, 1984,71(6):873-877.
[14]	刘军, 王小伟, 魏钦平, 鲁韧强, 高照全 . 世界葡萄抗寒育种的成就与展望. 果树学报, 2004,21(5):461-466. doi: 10.3969/j.issn.1009-9980.2004.05.016
	LIU J, WANG X W, WEI Q P, LU R Q, GAO Z Q . Achievements and prospect of world cold-resistant grape breeding. Journal of Fruit Science, 2004,21(5):461-466. (in Chinese) doi: 10.3969/j.issn.1009-9980.2004.05.016
[15]	屈田田 . 葡萄胚挽救育种应用研究[D]. 杨凌: 西北农林科技大学, 2016.
	QU T T . Study on the application of grape breeding using embryo rescue[D]. Yangling: Northwest A&F University, 2016. (in Chinese)
[16]	张剑侠, 牛茹萱 . 无核葡萄胚挽救技术的研究现状与展望. 园艺学报, 2013,40(9):1645-1655.
	ZHANG J X, NIU R X . The present situation and prospect of embryo rescue technique research in seedless grape breeding. Acta Horticulturae Sinica, 2013,40(9):1645-1655. (in Chinese)
[17]	孟新法, 张利, 张潞生, 张子莲 . 无核葡萄胚发育及早期离体培养的研究: Ⅲ. 胚珠接种时期对胚发育的影响. 北京农业大学学报, 1993,19(1):45-47.
	MENG X F, ZHANG L, ZHANG L S, ZHANG Z L . A study on ovule development of seedless grapes and its early culture in vitro: Ⅲ. The effect of inoculation dates on embryo development. Acta Agriculturae Universitatis Pekinensis, 1993,19(1):45-47. (in Chinese)
[18]	徐海英, 张国军, 闫爱玲 . 无核葡萄育种及杂交亲本的选择. 中外葡萄与葡萄酒, 2001(3):30-32. doi: 10.3969/j.issn.1004-7360.2001.03.011
	XU H Y, ZHANG G J, YAN A L . The seedless grape breeding and parental selection. Sino-Overseas Grapevine and Wine, 2001(3):30-32. (in Chinese) doi: 10.3969/j.issn.1004-7360.2001.03.011
[19]	GARCIA E, MARTINEZ A, GARCIA DE LA CALERA E, PEREA L J, CENIS J L, CARRENO J . In vitro culture of ovules and embryos of grape for the obtention of new seedless table grape cultivars. Acta Horticulturae, 2000,528:663-666. doi: 10.17660/ActaHortic.2000.528.99
[20]	田莉莉 . 抗病无核葡萄胚挽救育种及种质创新[D]. 杨凌: 西北农林科技大学, 2007.
	TIAN L L . Breeding for the disease-resistant seedless grape novel varieties and innovation of new germplasms using embryo rescue[D]. Yangling: Northwest A&F University, 2007. ( in Chinese)
[21]	赵月玲, 程贯召, 王汉海 . 无核葡萄的育种与胚挽救技术的运用. 生物学通报, 2004,39(9):15-16. doi: 10.3969/j.issn.0006-3193.2004.09.007
	ZHAO Y L, CHENG G Z, WANG H H . Seedless grape breeding and application of embryo rescue technology. Bulletin of Biology, 2004,39(9):15-16. (in Chinese) doi: 10.3969/j.issn.0006-3193.2004.09.007
[22]	PONCE M T, AGUERO C B, GREGORI M T, TIZIO R . Factors affecting the development of stenospermic grape (Vitis vinifera) embryos cultured in vitro. Acta Horticulturae, 2000,528:667-672.
[23]	唐冬梅, 蔡军社, 骆强伟, 廖新福, 孙锋, 赵荣华, 符晓敏 . 用于无核葡萄选育的胚挽救技术研究. 果树学报, 2008,25(3):316-321. doi: 10.3969/j.issn.1009-9980.2008.03.006
	TANG D M, CAI J S, LUO Q W, LIAO X F, SUN F, ZHAO R H, FU X M . Study on embryo rescue technique for seedless grape breeding. Journal of Fruit Science, 2008,25(3):316-321. (in Chinese) doi: 10.3969/j.issn.1009-9980.2008.03.006
[24]	温晓敏, 张娜, 路凤珍, 田淑芬 . 红宝石无核和希姆劳特葡萄杂交胚挽救技术研究. 天津师范大学学报(自然科学版), 2017,37(5):32-35.
	WEN X M, ZHANG N, LU F Z, TIAN S F . Embryo rescue techniques of the reciprocal cross between Ruby Seedless grape and Himrod Seedless grape. Journal of Tianjin Normal University (Nature Science Edition), 2017,37(5):32-35. (in Chinese)
[25]	宋润刚, 路文鹏, 沈育杰, 金仁浩, 李晓红, 郭振贵, 刘景宽, 林兴桂 . 酿酒葡萄新品种‘北冰红’. 园艺学报, 2008,35(7):1085. doi: 10.3321/j.issn:0513-353X.2008.07.024
	SONG R G, LU W P, SHEN Y J, JIN R H, LI X H, GUO Z G, LIU J K, LIN X G . A new ice-red brewing grape cultivar ‘Beibinghong’. Acta Horticulturae Sinica, 2008,35(7):1085. (in Chinese) doi: 10.3321/j.issn:0513-353X.2008.07.024
[26]	皇甫淳, 张辉, 修荆昌, 冯玉才 . “双优”两性花山葡萄新品种选育研究. 中外葡萄与葡萄酒, 1994(4):51-53.
	HUANGFU C, ZHANG H, XIU J C, FENG Y C . A study on selection of a new breeding of hermaphrodite grape ‘Shuangyou’.Sino- Overseas Grapevine and Wine, 1994(4):51-53. (in Chinese)
[27]	黎盛臣, 文丽珠, 张凤琴, 罗芳梅, 杨美容, 张映祝, 黄德藩 . 抗寒抗病葡萄新品种——北醇. 植物学报, 1983,1(2):28-30.
	LI S C, WEN L Z, ZHANG F Q, LUO F M, YANG M R, ZHANG Y Z, HUANG D F . A new grape variety for cold-resistance and disease-resistance——Beichun. Chinese Bulletin of Botany, 1983,1(2):28-30. (in Chinese)
[28]	王爱玲, 王跃进, 唐冬梅, 张剑侠, 张朝红 . 提高无核葡萄胚挽救中幼胚成苗率的研究. 中国农业科学, 2010,43(20):4238-4245.
	WANG A L, WANG Y J, TANG D M, ZHANG J X, ZHANG C H . Research on improvement of seedling rate in embryo rescue of seedless grapes. Scientia Agricultura Sinica, 2010,43(20):4238-4245. (in Chinese)
[29]	LAHOGUE F, THIS P, BOUQUET A . Identification of a codominant scar marker linked to the seedlessness character in grapevine. Theoretical and Applied Genetics, 1998,97(5/6):950-959. doi: 10.1007/s001220050976
[30]	MEHIA N, HINRICHSEN P . A new, highly assertive scar marker potentially useful to assist selection for seedlessness in table grape breeding. Acta Horticulturae, 2003,603:559-564. doi: 10.17660/ActaHortic.2003.603.74
[31]	王跃进, 杨英军, 周鹏, 张剑侠, 王西平 . 用DNA探针检测我国栽培的无核葡萄及辅助育种初探. 园艺学报, 2002,29(2):105-108. doi: 10.3321/j.issn:0513-353X.2002.02.003
	WANG Y J, YANG Y J, ZHOU P, ZHANG J X, WANG X P . Detecting the seedless characteristics of the grapes in China with DNA probe and DNA marker assistant selection. Acta Horticulturae Sinica, 2002,29(2):105-108. (in Chinese) doi: 10.3321/j.issn:0513-353X.2002.02.003
[32]	CABEZAS J A, CERVERA M T, RUIZ-GARCIA L, CARREñO J, MARTíNEZ-ZAPATER J M . A genetic analysis of seed and berry weight in grapevine. Genome, 2006,49(12):1572-1585. doi: 10.1139/g06-122
[33]	MEJIA N, SOTO B, GUERRERO M, CASANUEVA X, HOUEL C, MICCONO MDE L, RAMOS R, LE CUNFF L, BOURSIQUOT J M, HINRICHSER P， ADAM-BLONDON A F . Molecular, genetic and transcriptional evidence for a role of VvAGL11 in stenospermocarpic seedlessness in grapevine. BMC Plant Biology, 2011,11:57. doi: 10.1186/1471-2229-11-57 pmid: 3076230

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

杂交组合 Cross combination	母本Female parent		父本Male parent
杂交组合 Cross combination	种或杂种 Species or hybrid	特性 Characteristic	种或杂种 Species or hybrid	特性 Characteristic
昆香无核×双优 Kunxiang Seedless × Shuangyou	欧亚种 V. vinifera	无核、玫瑰香味 Seedless, aroma of muscat	山葡萄 V. amurensis	有核、抗寒 Seeded, cold-resistance
无核白×双优 Thompson Seedles × Shuangyou	欧亚种 V. vinifera	无核 Seedless	山葡萄 V. amurensis	有核、抗寒 Seeded, cold-resistance
红脸无核×双优 Blush Seedless × Shuangyou	欧亚种 V. vinifera	无核 Seedless	山葡萄 V. amurensis	有核、抗寒 Seeded, cold-resistance
美丽无核×北冰红 Beauty Seedless × Beibinghong	欧亚种 V. vinifera	无核 Seedless	欧山杂种 V. amurensis × (V. amurensis × V. vinifera)	有核、抗寒 Seeded, cold-resistance
无核白×北冰红 Thompson Seedless × Beibinghong	欧亚种 V. vinifera	无核 Seedless	欧山杂种 V. amurensis × (V. amurensis × V. vinifera)	有核、抗寒 Seeded, cold-resistance
红脸无核×北冰红 Blush Seedless × Beibinghong	欧亚种 V. vinifera	无核 Seedless	欧山杂种 V. amurensis × (V. amurensis × V. vinifera)	有核、抗寒 Seeded, cold-resistance
红宝石无核×北醇 Ruby Seedless × Beichun	欧亚种 V. vinifera	无核 Seedless	欧山杂种 V. vinifera × V. amurensis	有核、抗寒、抗病 Seeded, cold-resistance, disease-resistance

杂交组合 Cross combination	授粉时间 Pollination time	取样时间Sampling time	接种胚珠数Number of ovules	发育胚Embryos developed		胚萌发成苗Plantlet formation
杂交组合 Cross combination	授粉时间 Pollination time	取样时间Sampling time	接种胚珠数Number of ovules	Number	%	Number	%
昆香无核×双优 Kunxiang Seedless × Shuangyou	05-15	06-27	141	64	45.39	23	16.31
无核白×双优 Thompson Seedless × Shuangyou	05-19	06-23	135	26	19.26	11	8.15
红脸无核×双优 Blush Seedless × Shuangyou	05-17	07-13	200	78	39.00	19	9.50
美丽无核×北冰红 Beauty Seedless × Beibinghong	05-17	06-23	203	21	10.34	3	1.48
无核白×北冰红 Thompson Seedless × Beibinghong	05-19	06-23	157	12	7.64	3	1.91
红脸无核×北冰红 Blush Seedless × Beibinghong	05-17	07-13	182	61	33.52	12	6.59
红宝石无核×北醇 Ruby Seedless × Beichun	05-18	07-13	150	69	46.00	26	17.33
∑			1168	331	—	97	—

杂交组合 Cross combination	培养基 Culture medium		胚珠数 Number of ovule	发育胚 Embryos developed		胚萌发成苗 Plantlet formation
杂交组合 Cross combination	培养基 Culture medium		胚珠数 Number of ovule	Number	%	Number	%
无核白×双优 Thompson Seedless × Shuangyou	基本培养基 Basal medium	MM3	135	26	19.26a	11	8.15a
无核白×双优 Thompson Seedless × Shuangyou		ER	195	26	13.33b	7	3.59b
红宝石无核×北醇 Ruby Seedless × Beichun		MM3	150	69	46.00a	26	17.33a
红宝石无核×北醇 Ruby Seedless × Beichun		ER	120	42	35.00b	11	9.17b
昆香无核×双优 Kunxiang Seedless × Shuangyou	添加不同氨基酸的培养基 Medium with different amino acids	MM3+2.5 mmol·L^-1半胱氨酸Cysteine	150	62	41.33c	28	18.67ab
		MM3+2.5 mmol·L^-1天冬酰胺Asparagine	140	67	47.86bc	30	21.43a
		MM3+2.5 mmol·L^-1谷氨酰胺Glutamine	129	77	59.69a	27	20.93a
		MM3+2.5 mmol·L^-1丝氨酸Serine	165	88	53.33ab	22	13.33c
		MM3	141	64	45.39bc	23	16.31bc
红宝石无核×北醇 Ruby Seedless × Beichun		MM3+2.5 mmol·L^-1半胱氨酸Cysteine	435	205	47.13b	50	11.49c
		MM3+2.5 mmol·L^-1天冬酰胺Asparagine	70	39	55.71a	15	21.43a
		MM3+2.5 mmol·L^-1谷氨酰胺Glutamine	360	165	45.83b	80	22.22a
		MM3+2.5 mmol·L^-1丝氨酸Serine	340	151	44.41b	64	18.82b
		MM3	150	69	46.00b	26	17.33b

Breeding for Grape Germplasm Involved in Seedlessness with Cold-Resistant Using Embryo Rescue

RichHTML

PDF

Abstract

Cite this article

share this article

Figures/Tables 11

References 33

Related Articles 5

Metrics

Comments

Recommended 0

[1]	PeiPei ZHU,YiJia LUO,Wen XIANG,MingLei ZHANG,JianXia ZHANG. Rescue and Molecular Marker Assisted-Selection of the Cold-Resistant Seedless Grape Hybrid Embryo [J]. Scientia Agricultura Sinica, 2021, 54(6): 1218-1228.
[2]	LI GuiRong,QUAN Ran,CHENG ShanShan,HOU XiaoJin,FAN XiuCai,HU HuiLing. The Influencing Factors of in-vitro Ovule Development in Seedless Grape and Its Physiological Changes [J]. Scientia Agricultura Sinica, 2020, 53(22): 4646-4657.
[3]	WU Meng-jing, XU Qing-ye, LIU Ya, SHI Xing-rong, SHEN Qi-da, YANG Meng-meng, WANG Shi-gui, TANG Bin. The Super Cooling Point Change of Harmonia axyridis Under Low Temperature Stress and Its Cold-Resistance Genes’ Expression Analysis [J]. Scientia Agricultura Sinica, 2016, 49(4): 677-685.
[4]	WANG Ai-ling,WANG Yue-jin,TANG Dong-mei,ZHANG Jian-xia,ZHANG Chao-hong . Research on Improvement of Seedling Rate in Embryo Rescue of Seedless Grapes [J]. Scientia Agricultura Sinica, 2010, 43(20): 4238-4245 .
[5]	TANG Dong-mei,WANG Yue-jin,ZHAO Rong-hua,PAN Xue-jun,CAI Jun-she,ZHANG Jian-xia,ZHANG Chao-hong,LUO Qiang-wei . Factors Influencing Embryo Development in Embryo Rescue of Seedless Grapes#br# [J]. Scientia Agricultura Sinica, 2009, 42(7): 2449-2457 .