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作物学报 ›› 2020, Vol. 46 ›› Issue (3): 317-329.doi: 10.3724/SP.J.1006.2020.92036

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

水稻光温敏核不育基因tms5与pms3的互作效应

王芳权1,2,范方军1,2,夏士健1,宗寿余1,郑天清3,王军1,2,李文奇1,2,许扬1,2,陈智慧1,2,蒋彦婕1,2,陶亚军1,2,仲维功1,2,杨杰1,2,*()   

  1. 1. 江苏省农业科学院粮食作物研究所/国家水稻改良中心南京分中心/江苏省优质水稻工程技术研究中心, 江苏南京 210014
    2. 扬州大学江苏省粮食作物现代产业技术协同创新中心, 江苏扬州 225009
    3. 中国农业科学院作物科学研究所, 北京 100081
  • 收稿日期:2019-07-08 接受日期:2019-09-26 出版日期:2020-03-12 网络出版日期:2019-10-14
  • 通讯作者: 杨杰
  • 作者简介:E-mail: wfqjaas@163.com, Tel: 025-84390320
  • 基金资助:
    本研究由国家转基因生物新品种培育科技重大专项(2018ZX08001-02B);江苏省自然科学基金面上项目(BK20171326);江苏省现代农业重点研发项目资助(BE2018388);江苏省现代农业重点研发项目资助(BE2017368)

Interactive effects of the photoperiod-/thermo-sensitive genic male sterile genes tms5 and pms3 in rice

Fang-Quan WANG1,2,Fang-Jun FAN1,2,Shi-Jian XIA1,Shou-Yu ZONG1,Tian-Qing ZHENG3,Jun WANG1,2,Wen-Qi LI1,2,Yang XU1,2,Zhi-Hui CHEN1,2,Yan-Jie JIANG1,2,Ya-Jun TAO1,2,Wei-Gong ZHONG1,2,Jie YANG1,2,*()   

  1. 1. Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Nanjing Branch of Chinese National Center for Rice Improvement/Jiangsu High Quality Rice R&D Center, Nanjing 210014, Jiangsu, China;
    2. Jiangsu Co-innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, Jiangsu, China
    3. Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2019-07-08 Accepted:2019-09-26 Published:2020-03-12 Published online:2019-10-14
  • Contact: Jie YANG
  • Supported by:
    This study was supported by the National Major Project for Developing New GM Crops(2018ZX08001-02B);the Jiangsu Province Natural Science Foundation(BK20171326);and the Jiangsu Province Key Research and Development Program (Modern Agriculture)(BE2018388);and the Jiangsu Province Key Research and Development Program (Modern Agriculture)(BE2017368)

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摘要：

tms5与pms3是水稻的光温敏核不育基因, 其功能位点已经明确, 然而它们在两系不育系中的效应尚不清楚。本研究针对tms5与pms3基因功能位点, 分别设计了功能标记AS-TMS5和CAPS-PMS3。经鉴定发现, 这2个功能标记能准确区分不育、可育性状对应的隐性纯合、杂合和显性纯合3种基因型。利用AS-TMS5和CAPS-PMS3对培矮64S/9311、广占63S/湘恢47和粤光S/宁恢108的F2群体单株的基因型及育性的关系分析发现, tms5基因是广占63S和粤光S控制光温敏不育性状的主效基因, 而pms3基因在培矮64S和粤光S中并不能独立起作用, 还需要与其他基因共同调控。进一步分析粤光S/宁恢108的F2:3群体基因型与育性的关系, 发现在粤光S/宁恢108背景下, 携带pms3基因的株系几乎都表现可育, 而携带tms5基因的株系在较高气温条件下表现不育, 但育性转换温度可能较高; 而携带tms5与pms3基因的株系育性转换温度比仅携带tms5基因的株系低, 这为聚合2个基因选育不育性状稳定的光温敏不育系提供了思路和方法。

关键词: tms5, pms3, 功能标记, 光敏不育, 温敏不育

Abstract:

The tms5 and pms3 are two photoperiod-/thermo-sensitive genic male sterile genes, and their functional sites are clarified. However, the interactive effects of tms5 and pms3 in two-line sterile lines are still unclear. In this study, the functional markers AS-TMS5 and CAPS-PMS3 were designed, according to the functional sites of tms5 and pms3 respectively. The three genotypes of tms5 and pms3 were accurately distinguished by AS-TMS5 and CAPS-PMS3. The relationship of the phenotype and genotype in the F2 population of Pei’ai 64S/9311, Guangzhan 63S/Xianghui 47 and Yueguang S/Ninghui 108 were analyzed respectively. The tms5 was the major gene in Guangzhan 63S and Yueguang S, while pms3 was a non-independent gene in Pei’ai 64S and Yueguang S. By the phenotype and genotype analysis of the F2:3 population of Yueguang S/Ninghui 108, the plants carrying pms3 almost were fertile, while the plants carrying tms5 showed sterility, and had higher transition temperature. Furthermore, the sterility of the plants carrying tms5 and pms3 might have lower transition temperature than those carrying tms5. Pyramiding of tms5 and pms3 provides an efficient scheme to breed photoperiod-/thermo-sensitive genic male sterile lines, which have lower transition temperature and safer production.

Key words: tms5, pms3, functional marker, photoperiod-sensitive male sterility, thermo-sensitive male sterility

表1

TMS5、PMS3功能标记引物"

分子标记
Molecular marker		 引物名称
Primer name		 引物序列
Primer sequence (5′-3′)		 产物长度
Product length (bp)
AS-TMS5 T5n-F CCACCGGGTCGGCCGAAGGC 302
T5m-F CCACCGGGTCGGCCGAAGTA
T5-R AGCTCGAAGAGGCGCTCCAC
CAPS-PMS3 PMS3-F GCTTTCCCAGGATGCACATA 410
PMS3-R GTTTGCTCCATTGGTTAGGC

图1

2015年8月至9月田间气温走势图"

图2

水稻品种(系)的AS-TMS5和CAPS-PMS3标记检测 1~24分别为: 日本晴、9311、南京16号、湘恢47、宁恢108、农垦58S、安农S-1、株1S、培矮64S、广占63S、粤光S、N111S、C815S、Y58S、509S、武香S、深08S、1206S、1208S、L126S、L128S、丰39S、1892S和L816S。"

图3

TMS5基因突变位点的测序分析 (a)和(b)的差异位点位于TMS5基因编码区之前, (c)为起始密码子, (d)为功能突变位点。"

图4

PMS3基因突变位点的测序分析"

表2

粤光S/宁恢108 F2群体基因型与育性的关系"

编号
No.		 AS-
TMS5		 CAPS-
PMS3		 表型
Phenotype		 编号
No.		 AS-
TMS5		 CAPS-
PMS3		 表型
Phenotype		 编号
No.		 AS-
TMS5		 CAPS-
PMS3		 表型
Phenotype
Yueguang S + + S 26 * * F 54 - * F
Ninghui 108 - - F 27 * * F 55 * * F
F1 * * F 28 * - F 56 * * F
1 + * S 29 * * F 57 * + F
2 + * S 30 - + F 58 * * F
3 + * S 31 * * F 59 * * F
4 + * S 32 * * F 60 * * F
5 + + S 33 * - F 61 * + F
6 + * S 34 - - F 62 * + F
7 + * S 35 * - F 63 * * F
8 + - S 36 * * F 64 - * F
9 + * S 37 * * F 65 * - F
10 + * S 38 * * F 66 * + F
11 + * S 39 - * F 67 * + F
12 + + S 40 * * F 68 * * F
13 + + S 41 * - F 69 * + F
14 + - S 42 * - F 70 * - F
15 + * S 43 * * F 71 - + F
16 + + S 44 * * F 72 * - F
17 + - S 45 - + F 73 * * F
18 + + S 46 * + F 74 - - F
19 + * S 47 * - F 75 - * F
20 + * S 48 - - F 76 * + F
21 + * S 49 * * F 77 * - F
22 + * S 50 - * F 78 - * F
23 * - F 51 * - F 79 - - F
24 * - F 52 * - F
25 * * F 53 * * F

图5

以AS-TMS5和CAPS-PMS3标记检测粤光S/宁恢108 F2群体部分单株 P1: 粤光S; P2: 宁恢108; F1: 粤光S/宁恢108杂交种; 1~45: 粤光S/宁恢108 F2群体部分单株。"

表3

粤光S/宁恢108组合F2:3株系育性分析"

株系
Line		 基因型
Genotype		 第1期 Phase I 第2期 Phase II
抽穗期
Heading date
(month/day)		 总数
Total number		 不育株
No. of sterile plants		 可育株
No. of fertile plants		 抽穗期
Heading date
(month/day)		 总数
Total
number		 不育株
No. of sterile plants		 可育株
No. of fertile plants
S1 TTPP 8/26 11 0 11 9/3 10 0 10
S2 TTPP 8/27 11 0 11 9/9 10 0 10
S3 TTPP 9/11 11 0 11 9/25 10 0 10
S4 TTpp 8/28 11 0 11 9/3 10 0 10
S5 TTpp 9/5 11 0 11 9/14 11 0 11
S6 TTpp 8/20 11 0 11 9/9 11 0 11
S7 TTpp 9/7 11 0 11 9/19 11 0 11
S8 TTpp 9/7 11 1 10 9/22 11 0 11
S9 ttPP 8/28 11 11 0 9/16 11 6 5
S10 ttPP 8/30 11 11 0 9/17 11 2 9
S11 ttPP 9/1 11 11 0 9/17 10 5 5
S12 ttpp 8/23 11 11 0 9/15 10 9 1
S13 ttpp 8/24 11 11 0 9/10 10 10 0
S14 ttpp 8/30 11 11 0 9/13 11 11 0
S15 ttpp 9/10 10 10 0 9/19 9 9 0

图6

粤光S/宁恢108组合F2:3株系育性分析 A: 花药形态和花粉育性分析, 大图标尺为1 mm, 小图标尺为100 μm; B: 小穗结实分析, 标尺为2 cm。"

附表1

培矮64S/9311 F2群体基因型与育性的关系"

编号
No.		 CAPS-
PMS3		 表型
Phenotype		 编号
No.		 CAPS-
PMS3		 表型
Phenotype		 编号
No.		 CAPS-
PMS3		 表型
Phenotype
培矮64S Pei’ai 64S + S 20 * F 42 + F
9311 - F 21 * F 43 + F
F1 * F 22 + F 44 + F
1 + S 23 - F 45 * F
2 * S 24 * F 46 * F
3 + S 25 * F 47 + F
4 + S 26 * F 48 + F
5 * S 27 + F 49 * F
6 + S 28 * F 50 * F
7 * S 29 - F 51 - F
8 + S 30 * F 52 - F
9 + S 31 - F 53 * F
10 * F 32 - F 54 * F
11 - F 33 * F 55 * F

附表1

培矮64S/9311 F2群体基因型与育性的关系(续附表1)"

编号
No.		 CAPS-
PMS3		 表型
Phenotype		 编号
No.		 CAPS-
PMS3		 表型
Phenotype		 编号
No.		 CAPS-
PMS3		 表型
Phenotype
12 * F 34 * F 56 * F
13 + F 35 * F 57 * F
14 + F 36 - F 58 + F
15 * F 37 * F 59 - F
16 * F 38 + F 60 - F
17 + F 39 * F 61 * F
18 - F 40 - F 62 + F
19 + F 41 * F

附表2

广占63S/湘恢47 F2群体基因型与育性的关系"

编号
No.		 AS-
TMS5		 表型
Phenotype		 编号
No.		 AS-
TMS5		 表型
Phenotype		 编号
No.		 AS-
TMS5		 表型
Phenotype
广占63S Guangzhan 63S + S 29 + S 60 * F
湘恢47 Xianghui 47 - F 30 * F 61 * F
F1 * F 31 - F 62 * F
1 + S 32 * F 63 * F
2 + S 33 * F 64 * F
3 + S 34 - F 65 * F
4 + S 35 * F 66 - F
5 + S 36 - F 67 * F
6 + S 37 * F 68 * F
7 + S 38 - F 69 * F
8 + S 39 * F 70 - F
9 + S 40 * F 71 * F
10 + S 41 * F 72 * F
11 + S 42 * F 73 * F
12 + S 43 * F 74 - F
13 + S 44 * F 75 - F
14 + S 45 * F 76 * F
15 + S 46 * F 77 - F
16 + S 47 * F 78 * F
17 - S 48 * F 79 * F
18 + S 49 * F 80 - F
19 + S 50 * F 81 * F
20 + S 51 * F 82 * F
21 + S 52 * F 83 * F
22 + S 53 * F 84 * F
23 + S 54 - F 85 * F
24 + S 55 * F 86 - F
25 + S 56 - F 87 * F
26 + S 57 * F 88 - F
27 + S 58 * F 89 * F
28 + S 59 - F

附图1

2015年8月至9月田间日照走势图"

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[1] 陈日荣,周延彪,王黛君,赵新辉,唐晓丹,许世冲,唐倩莹,符星学,王凯,刘选明,杨远柱. 利用CRISPR/Cas9技术编辑水稻温敏不育基因TMS5[J]. 作物学报, 2020, 46(8): 1157-1165.
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