分子标记辅助选择改良C418对红莲型粳稻不育系的恢复力

doi:10.16819/j.1001-7216.2018.7152

摘要/Abstract

摘要：

【目的】高效选育红莲型(Honglian,HL)粳稻恢复系有助于HL型杂交粳稻育种,对促进三系杂交粳稻的发展具有重要的意义。【方法】Rf6是一个HL型恢复基因,来源于HL型籼稻强恢复系9311。前期研究中,在以9311为供体、日本晴为受体的一套染色体片段代换系中鉴定出携带Rf6的株系R1093。本研究利用R1093与BT型粳稻恢复系C418(携带Rf1)杂交,通过常规回交育种结合分子标记辅助选择技术,将Rf6导入C418中,进行Rf6与Rf1聚合育种;利用BT型、HL型六千辛A进行测交鉴定改良系的恢复力。【结果】共获得12个BC₃F₄株系和55个BC₄F₃株系,其中6个改良系的农艺性状已基本接近C418;测交鉴定结果表明聚合Rf6的改良系对HL型粳稻不育系的恢复度达到85%以上,可应用于水稻生产;对BT型粳稻不育系的恢复度提升效果不显著。【结论】聚合Rf6能有效改良BT型粳稻恢复系对HL型粳稻不育系恢复力,是选育HL型粳稻恢复系的一条重要途径。

关键词: 粳稻, 红莲型, 恢复系, 分子标记辅助选择

Abstract:

【Objective】Efficient breeding of japonica restorers for Honglian(HL) type japonica cytoplasmic male sterile(CMS) lines is vital to develop HL-type japonica hybrids, which is of great significance for the development of three-line japonica hybrids.【Method】Rf6, a fertility restorer gene for HL-type CMS, was identified from ‘9311’, an indica restorer for HL-type CMS lines. In our previous study, R1093, a chromosome segment substitution line derived from the cross between the donor parent ‘9311’and the recipient parent ‘Nipponbare’, had been identified to carry Rf6. In the present study, Rf6 was pyramided into C418, a BT-type japonica restorer only carrying Rf1, using backcross via marker-assisted selection, and the restoration ability of the improved lines for BT-, HL-type CMS lines was analyzed by testcrossing.【Result】To 2016, a total of 67 improved lines, including 12 BC₃F₄ lines and 55 BC₄F₃ lines, had been obtained, and most of the agronomic traits were well maintained in six improved lines. The testcrossed F₁ plants from improved lines and BT-Liuqianxin A exhibited the similar fertility levels with those of the testcrossed F₁ plants from C418. The significant increased fertility levels were observed on the testcrossed F₁ plants from improved lines and HL-Liuqianxin A, and most of these testcrossing lines exhibited fertility levels over 85%, indicating these improved lines can be applied to rice production.【Conclusions】The pyramiding of Rf6 and Rf1 can improve the restoration ability of BT-type japonica restorer lines to HL-type japonica CMS lines, which should be an important way to breed HL-type japonica restorer lines.

Key words: japonica, Honglian-type, restorer line, marker-assisted selection

中图分类号:

张宏根, 仲崇元, 司华, 刘巧泉, 顾铭洪, 汤述翥. 分子标记辅助选择改良C418对红莲型粳稻不育系的恢复力[J]. 中国水稻科学, 2018, 32(5): 445-452.

Honggen ZHANG, Chongyuan ZHONG, Hua SI, Qiaoquan LIU, Minghong GU, Shuzhu TANG. Improving the Ability of C418 to Restore the Fertility of the Honglian-type Cytoplasmic Male Sterility japonica Lines via Molecular Marker-assisted Selection[J]. Chinese Journal OF Rice Science, 2018, 32(5): 445-452.

图/表 8

参考文献 21

[1]	邓华凤, 何强, 舒服, 张武汉, 杨飞, 荆彦辉, 东丽, 谢辉. 中国杂交粳稻研究现状与对策. 杂交水稻, 2006, 21(1): 1-6.
	Deng H F, He Q, Shu F, Zhang W H, Yang F, Jing Y H, Dong L, Xie H.Status and technical strategy on development of japonica hybrid rice in China. Hybrid Rice, 2006, 21(1): 1-6. (in Chinese with English abstract)
[2]	Fujimura T, Akagi H, Oka M, Nakamura A, Sawada R.Establishment of a rice protoplast culture and application of an asymmetric protoplast fusion technique to hybrid rice breeding. J Food Sci, 2010, 13(3): 243-247.
[3]	曾千春, 周开达, 朱祯, 罗琼. 中国水稻杂种优势利用现状. 中国水稻科学, 2000, 14(4): 243-246.
	Zeng Q C, Zhou K D, Zhu Z, Luo Q.Current status in the use of hybrid rice heterosis in China.Chin J Rice Sci, 2000, 14(4): 243-246. (in Chinese with English abstract)
[4]	汤述翥, 张宏根, 梁国华, 严长杰, 刘巧泉, 顾铭洪. 三系杂交粳稻发展缓慢的原因及对策. 杂交水稻, 2008, 23(1): 1-5.
	Tang S Z, Zhang H G, Liang G H, Yan C J, Liu Q Q, Gu M H.Reasons and counter measures of slow development on three-line japonica hybrid rice. Hybrid Rice, 2008, 23(1): 1-5. (in Chinese with English abstract)
[5]	汤述翥, 孙叶, 张宏根, 顾燕娟, 陆驹飞, 田舜, 余波, 顾铭洪. 同核异质粳稻不育系特性比较. 中国水稻科学2005, 19(6): 521-526.
	Tang S Z, Sun Y, Zhang H G, Gu Y J, Lu J F, Tian S, Yu B, Gu M H.Comparison on the characteristics of the isonuclear alloplasmic CMS lines in japonica rice. Chin J Rice Sci, 2005, 19(6): 521-526. (in Chinese with English abstract).
[6]	汤述翥, 张宏根, 朱正斌, 刘超, 李鹏, 梁国华, 严长杰, 刘巧泉, 于恒秀, 顾铭洪. 红莲型不育细胞质在杂交粳稻育种中的应用. 中国水稻科学, 2010, 24(2): 116-124.
	Tang S Z, Zhang H G, Zhu Z B, Liu C, Li P, Liang G H, Yan C J, Liu Q Q, Yu H X, Gu M H.Application of HL type male sterile cytoplasm in japonica hybrid rice breeding. Chin J Rice Sci, 2010, 24(2): 116-124. (in Chinese with English abstract)
[7]	汤述翥, 张宏根, 朱正斌, 刘巧泉, 梁国华, 严长杰, 刘超, 李鹏, 顾铭洪. 红莲型不育细胞质应用于粳稻杂种优势的思考与初探. 西南农业学报, 2009, 22(4): 1158-1164.
	Tang S Z, Zhang H G, Zhu Z B, Liu Q Q, Liang G H, Yan C J, Liu C, Li P, Gu M H.Toward to apply HL-type male sterile cytoplasm for japonica heterosis utilization: Consideration and practice. Southwest China J Agric Sci, 2009, 22(4): 1158-1164. (in Chinese with English abstract)
[8]	朱正斌, 张宏根, 刘超, 李鹏, 裔传灯, 汤述翥, 顾铭洪. 四种细胞质六千辛A粳稻不育系育种利用特性的比较研究. 作物学报, 2010, 36(1): 1-8.
	Zhu Z B, Zhang H G, Liu C, Li P, Yi C D, Tang S Z, Gu M H.Comparative study on breeding utilization characteristics of the isonuclear alloplasmic japonica CMS lines Liuqianxin A with four different cytoplasm sources. Acta Agron Sin, 2010, 36(1): 1-8. (in Chinese with English abstract)
[9]	Wang Z, Zou Y, Li X, Zhang Q, Chen L, Wu H, Su D, Chen Y, Guo J, Luo D, Long Y, Zhong Y, Liu Y.Cytoplasmic male sterility of rice with Boro II cytoplasm is caused by a cytotoxic peptide and is restored by two related PPR motif genes via distinct modes of mRNA silencing.Plant Cell, 2006, 18(3): 676-687.
[10]	Akagi H, Nakamura A, Yokozeki-Misono Y, Inagaki A, Takahashi H, Mori K, Fujimura T.Positional cloning of the rice Rf-1 gene, a restorer of BT-type cytoplasmic male sterility that encodes a mitochondria-targeting PPR protein. Theor Appl Genet, 2004, 108(8): 1449-1457.
[11]	Huang Q, He Y, Jing R, Zhu R, Zhu Y.Mapping of the nuclear fertility restorer gene for HL cytoplasmic male sterility in rice using microsatellite markers.Chin Sci Bull, 2000, 45(5): 430-432.
[12]	Huang J, Hu J, Xu X, Li S, Yi P, Yang D, Ren F, Liu X, Zhu Y.Fine mapping of the nuclear fertility restorer gene for HL cytoplasmic male sterility in rice.Bot Bull Acad Sin, 2003, 44(4): 285-289.
[13]	Hu J, Wang K, Huang W, Liu G, Gao Y, Wang J, Huang Q, Ji Y, Qin X, Wan L, Zhu R, Li S, Yang D, Zhu Y. The rice pentatricopeptide repeat protein RF5 restores fertility in Hong-Lian cytoplasmic male-sterile lines via a complex with the glycine-rich protein GRP162. Plant Cell, 2012, 24(1): 109-122.
[14]	Huang W, Hu J, Yu C, Huang Q, Wan L, Wang L, Qin X, Ji Y, Zhu R, Li S, Zhu Y.Two non-allelic nuclear genes restore fertility in a gametophytic pattern and enhance abiotic stress tolerance in the hybrid rice plant. Theor Appl Genet, 2012, 124(5): 799-807.
[15]	Huang W, Yu C, Hu J, Wang L, Dan Z, Zhou W, He C, Zeng Y, Yao G, Qi J, Zhang Z, Zhu R, Chen X, Zhu Y.Pentatricopeptide-repeat family protein RF6 functions with hexokinase 6 to rescue rice cytoplasmic male sterility.Proc Natl Acad Sci, 2015, 112(48): 14984-14989.
[16]	Zhang H, Che J, Ge Y, Pei Y, Zhang L, Liu Q, Tang S, Gu M.Ability of Rf5 and Rf6 to restore fertility of Chinsurah Boro II-type cytoplasmic male sterile Oryza sativa(ssp. japonica) lines. Rice, 2017, 10(1): 2.
[17]	Rogers S O, Bendich A J.Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues. Plant Mol Biol, 1985, 5: 69-76.
[18]	Zhang H, Zhang L, Si H, Ge Y, Liang G, Gu M, Tang S.Rf5 is able to partially restore fertility to Honglian-type cytoplasmic male sterile japonica rice(Oryza sativa ) lines. Mol Breeding, 2016, 36(7): 102.
[19]	杨振玉, 陈秋柏, 陈荣芳, 苏正基, 贾宝清, 佟景兴, 王健群. 水稻粳型恢复系C57的选育. 作物学报, 1981, 7(3): 153-156.
	Yang Z Y, Chen Q B, Chen R F, Su Z J, Jia B Q, Tong J X, Wang J Q.The breeding of japonica rice restorer C57. Acta Agron Sin, 1981, 7(3): 153-156. (in Chinese with English abstract)
[20]	杨振玉, 张宗旭, 魏耀林, 赵迎春, 高勇. 粳型特异亲和恢复系C418的选育及其特性. 杂交水稻, 1998(3): 33-34.
	Yang Z Y, Zhang Z X, Wei Y L, Zhao Y C, Gao Y.Breeding and characteristics ofjaponica type wide compatibility line C418. Hybrid Rice, 1998(3): 33-34. (in Chinese with English abstract)
[21]	赵迎春, 杨振玉, 魏耀林, 张忠旭. 从C418的育成再论北方杂交粳稻的发展形势. 杂交水稻, 2000(5): 5-6.
	Zhao Y C, Yang Z Y, Wei Y L, Zhang Z X.A second discussion on development of northernjaponica hybrid rice from the successful breeding of restorer C418. Hybrid Rice, 2000(5): 5-6. (in Chinese with English abstract)

基因 Gene	标记 Marker	正向序列 Forward primer(5′→3′)	反向序列 Reverse primer(5′→3′)
Rf6	RM407	GACTACGAGACGAGTGATTTGAACC	GCGTGGGAAATGACTAGGAGTAGG
Rf1	STS10-16	CAGCAATCGGATCGCCTC	ATCTTTCCGTGATGGGAGGTA

基因 Gene	标记 Marker	正向序列 Forward primer(5′→3′)	反向序列 Reverse primer(5′→3′)
Rf6	RM407	GACTACGAGACGAGTGATTTGAACC	GCGTGGGAAATGACTAGGAGTAGG
Rf1	STS10-16	CAGCAATCGGATCGCCTC	ATCTTTCCGTGATGGGAGGTA

世代 Generation	植株数 Number of measured plants	基因型 Genotype			频率 Frequency/ %
世代 Generation	植株数 Number of measured plants	Rf6Rf6	Rf6rf6	rf6rf6	频率 Frequency/ %
BC₁F₁	160	0	75	85	46.9
BC₂F₁	100	0	47	53	47.0
BC₂F₂	120	29	54	37	24.2
BC₃F₁	19	0	9	10	47.3
BC₄F₁	298	0	124	174	41.6
BC₄F₂	1000	243	461	296	24.3

世代 Generation	植株数 Number of measured plants	基因型 Genotype			频率 Frequency/ %
世代 Generation	植株数 Number of measured plants	Rf6Rf6	Rf6rf6	rf6rf6	频率 Frequency/ %
BC₁F₁	160	0	75	85	46.9
BC₂F₁	100	0	47	53	47.0
BC₂F₂	120	29	54	37	24.2
BC₃F₁	19	0	9	10	47.3
BC₄F₁	298	0	124	174	41.6
BC₄F₂	1000	243	461	296	24.3

株系号 Line	基因型 Genotype	BT型六千辛A BT-type Liuqianxin A	HL型六千辛A HL-type Liuqianxin A
SL1	Rf1Rf1Rf6Rf6	91.03±4.98	95.09±1.69^**
SL2	Rf1Rf1Rf6Rf6	91.65±3.06	90.87±1.90^**
SL3	Rf1Rf1Rf6Rf6	92.84±3.72	92.25±3.42^**
SL4	Rf1Rf1Rf6Rf6	94.62±3.13	94.14±2.53^**
SL5	Rf1Rf1Rf6Rf6	95.82±1.75	91.49±3.91^**
SL6	Rf1Rf1Rf6Rf6	92.96±2.00	93.00±1.06^**
SL7	Rf1Rf1Rf6Rf6	97.13±2.02	85.89±7.25^**
SL8	Rf1Rf1Rf6Rf6	94.13±3.51	94.30±2.74^**
SL9	Rf1Rf1Rf6Rf6	94.18±2.25	95.02±2.83^**
SL10	Rf1Rf1Rf6Rf6	93.60±2.28	83.75±8.52^**
C418	Rf1Rf1rf6rf6	91.88±3.02	74.49±5.06