利用远缘杂交和未减数配子基因创制小麦加倍单倍体

doi:10.3724/SP.J.1006.2023.31009

作物学报 ›› 2023, Vol. 49 ›› Issue (12): 3154-3161.doi: 10.3724/SP.J.1006.2023.31009

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

利用远缘杂交和未减数配子基因创制小麦加倍单倍体

刘小娟¹^,²(), 柳欣², 张明虎², 郝明², 甯顺腙², 袁中伟², 黄林², 刘登才¹^,², 张连全¹^,²^,^*()

¹四川农业大学 / 西南作物基因资源发掘与利用国家重点实验室, 四川成都 611130
²四川农业大学小麦研究所, 四川成都611130

收稿日期:2023-02-08 接受日期:2023-05-24 出版日期:2023-12-12 网络出版日期:2023-06-02
通讯作者: * 张连全, E-mail: zhanglianquan1977@126.com
作者简介:E-mail: 925095898@qq.com
基金资助:
国家自然科学基金项目(31671682);四川省重点研发项目(2021YFYZ0002)

Creation of doubled haploid in wheat using distant hybridization and unreduced gamete genes

LIU Xiao-Juan¹^,²(), LIU Xin², ZHANG Ming-Hu², HAO Ming², NING Shun-Zong², YUAN Zhong-Wei², HUANG Lin², LIU Deng-Cai¹^,², ZHANG Lian-Quan¹^,²^,^*()

¹State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China / Sichuan Agricultural University, Chengdu 611130, Sichuan, China
²Triticeae Research Institute, Sichuan Agricultural University, Chengdu 611130, Sichuan, China

Received:2023-02-08 Accepted:2023-05-24 Published:2023-12-12 Published online:2023-06-02
Contact: * E-mail: zhanglianquan1977@126.com
Supported by:
National Natural Science Foundation of China(31671682);Key Research and Development Program of Sichuan Province, China(2021YFYZ0002)

摘要/Abstract

摘要：

未减数配子的结合实现染色体自动加倍, 是多倍体物种起源的重要途径, 也是提高作物单倍体育种效率的重要手段。我们前期从四倍体小麦发掘出控制未减数配子形成的强效QTL位点QTug.sau-3B, 并通过人工合成小麦为“桥梁”, 将其导入到综合农艺性状优良的小麦新品系中。本实验使用5份含未减数配子基因的优良小麦新品系与不含未减数配子基因的小麦推广品种的F₁杂种作母本与3份白茅(Imperata cylindrica)进行远缘杂交, 共授粉4610朵小花, 结实1965粒, 经幼胚拯救获得244个幼胚, 其中50个幼胚发育正常生长为50个小麦单倍体植株。由于小麦单倍体植株未减数配子基因的表达易受环境影响, 因此, 对单倍体植株在相同光周期(18 h光照/6 h黑暗)下进行了不同温度25℃/18℃、25℃/15℃和25℃/10℃处理, 结果表明, 25℃/18℃和25℃/10℃条件下编号为H31单倍体植株能够结实, 自交结实率分别为4.35%和2.41%。该研究结果为建立“基于小麦-白茅杂交实现染色体消除和未减数配子基因实现染色体自动加倍”的小麦单倍体育种技术提供了参考。

关键词: 单倍体育种, 白茅, 未减数配子, 幼胚拯救

Abstract:

The union of unreduced female and male gametes leads to spontaneous chromosome doubling, which is not only an important way of the origin of polyploid species but also an essential tool for enhancing crop doubled haploid breeding efficiency. A major quantitative trait locus QTug.sau-3B responsible for unreduced gametes formation was discovered in tetraploid Triticum turgidum wheat and was further transferred to elite common wheat lines using synthetic hexaploid wheat as a bridge. This objective of this study is to make wheat/wheat F₁ hybrids between the elite lines with this gene region and commercial cultivars and to evaluate the efficiency of QTug.sau-3B gene leading to spontaneous chromosome doubling. Wheat/wheat F₁ hybrids and their parents were pollinated with the fresh pollen of Imperata cylindrica to produce wheat haploid through chromosome elimination of I. cylindrica. Doubled haploid (DH) were then developed by spontaneous chromosome doubling in haploids because of the union of unreduced female and male gametes. In this experiment, 5 F₁ hybrid materials and 3 I. cylindrica materials were used for distant hybridization. 4610 florets were pollinated, 1965 seeds were produced, 244 embryos were obtained, and 50 wheat haploid plants were obtained. In different temperature treatments, the haploid plant numbered H31 set seeds at 25℃/18℃ (18 h/6 h) and 25℃/10℃ (18 h/6 h), and the self-fertilization rate was 4.35% and 2.41%, respectively. The results of this study provide a reference for the establishment of wheat haploid breeding technology of chromosome elimination based on wheat-I. cylindrica hybridization and automatic chromosome doubling based on unreduced gamete genes.

Key words: haploid breeding, Imperata cylindrica, unreduced gametes, embryo rescue

刘小娟, 柳欣, 张明虎, 郝明, 甯顺腙, 袁中伟, 黄林, 刘登才, 张连全. 利用远缘杂交和未减数配子基因创制小麦加倍单倍体[J]. 作物学报, 2023, 49(12): 3154-3161.

LIU Xiao-Juan, LIU Xin, ZHANG Ming-Hu, HAO Ming, NING Shun-Zong, YUAN Zhong-Wei, HUANG Lin, LIU Deng-Cai, ZHANG Lian-Quan. Creation of doubled haploid in wheat using distant hybridization and unreduced gamete genes[J]. Acta Agronomica Sinica, 2023, 49(12): 3154-3161.

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编号 Code	来源地 Location	抽穗时间 Heading time (month/day)	开花时间 Flowering time (month/day)
BMZ1616	四川宜宾翠屏 Cuiping, Yibin, Sichuan	4/23	4/28
BMZ1617	四川资阳乐至 Lezhi, Ziyang, Sichuan	4/26	4/29
BMZ1801	四川成都崇州 Chongzhou, Chengdu, Sichuan	4/26	4/29
BMZ1605	江苏宿迁沭阳 Shuyang, Suqian, Jiangsu	4/27	4/29
BMZ1602	福建宁德屏南 Pingnan, Ningde, Fujian	4/28	4/30
BMZ1612	山东临沂费县 Feixian, Linyi, Shandong	4/28	4/30
BMZ1604	广东揭阳东山 Dongshan, Jieyang, Guangdong	4/28	5/1
BMZ1610	湖北十堰丹江口 Danjiangkou, Shiyan, Hubei	4/29	5/1
BMZ1621	江苏徐州沛县Peixian, Xuzhou, Jiangsu	4/29	5/1
BMZ1606	贵州遵义习水 Xishui, Zunyi, Guizhou	5/6	5/8
BMZ1618	四川自贡富顺 Fushun, Zigong, Sichuan	5/6	5/8
BMZ1619	浙江温州永嘉 Yongjia, Wenzhou, Zhejiang	5/9	5/11

小麦材料 Wheat material	授粉小花数 Pollinated florets	结实数 Pseudo seeds (%)	得胚数 Embryo formation (%)	获得植株 Plantlet rate (%)
L14-6/CM602 F₁	1588	701 (44.14)^c	169 (24.11)^a	26 (15.38)^b
L14-6/SM114 F₁	868	439 (50.58)^b	61 (13.9)^b	21 (34.43)^a
L13-81/SM114 F₁	963	395 (41.02)^c	9 (2.28)^c	0 (0)
L13-471/CM96 F₁	973	299 (30.73)^d	5 (1.67)^c	3 (60)^a
L13-316/SM830 F₁	218	131 (60.09)^a	0 (0)	0 (0)
总计Total	4610	1965 (42.62)	244 (12.42)	50 (20.49)

白茅编号 I. cylindrica	授粉小花数 Pollinated florets	结实数 Pseudo seeds (%)	得胚数 Embryo formation (%)	获得植株 Plantlet rate (%)
BMZ1801	430	146 (33.95)^b	30 (20.55)^a	8 (26.67)^a
BMZ1617	1418	514 (36.25)^b	28 (5.45)^c	5 (17.86)^a
BMZ1616	2762	1305 (47.25)^a	186 (14.25)^b	37 (19.89)^a

含QTug.sau-3B单株编号 Code of haploid plants with QTug.sau-3B	单倍体株数(编号) Number of haploid plants (Plant ID)	系谱 Pedigree	QTug.sau-3B来源 Origin of QTug.sau-3B
H1	3 (H1-H3)	L13-471/CM96//I. cylindrica F₁	AS2255
H7, H8, H10, H11, H12, H13, H14, H16, H19, H20, H23, H24	21 (H4-H24)	L14-6/SM114//I. cylindrica F₁	Langdon
H25, H26, H27, H28, H31, H33, H35, H36, H38, H39, H41, H45, H47, H48, H49, H50	26 (H25-H50)	L14-6/CM602//I. cylindrica F₁	Langdon

单倍体编号 Haploid number	处理时期 Treatment stage	处理温度和光照时间 Temperature (℃)/time (h)	自交小花数 Selfed florets	自交结实数 Selfed seeds	自交结实率 Selfed seed set rate (%)
H31-1	孕穗期Booting stage	25/18, 18/6	92	4	4.35
H31-2	孕穗期Booting stage	25/15, 18/6	89	0	0
H31-3	孕穗期Booting stage	25/10, 18/6	83	2	2.41

利用远缘杂交和未减数配子基因创制小麦加倍单倍体

Creation of doubled haploid in wheat using distant hybridization and unreduced gamete genes

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材料 Material	株高 Plant height (cm)	分蘖数 No. of tillers	穗长 Spike length (cm)	小穗数 Spikelets	小花数 Florets	穗粒数 Seeds	自交结实率 Self-fertilization rate (%)
DH-H31-1	114.0	4	12.0	15	30	27	90.00
DH-H31-2	118.5	4	12.5	14	28	26	86.67
DH-H31-3	132.5	5	13.5	17	34	29	96.67
DH-H31-4	115.0	3	13.0	15	30	24	80.00

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